Azithromycin derivatives containing a phosphonium ion as anticancer agents

ABSTRACT

This invention relates to compounds that are useful as cancer therapies. The compounds comprise azithromycin derivatives having a phosphonium cation tethered to the azithromycin macrocycle. The invention also relates to methods of using said compounds and to pharmaceutical formulations comprising said compounds.

This invention relates to compounds that disrupt cell function, such asthe disruption of cell metabolism in particular cancer cell metabolism,that are useful as cancer therapies. The compounds comprise azithromycinderivatives having a phosphonium cation tethered to the azithromycinmacrocycle. The invention also relates to methods of using saidcompounds and to pharmaceutical formulations comprising said compounds.

BACKGROUND

Cancer is the fourth greatest cause of mortality in the developed world.In 2016 it was predicted that more than 1.6 million new cases of cancerwould be diagnosed in the U.S. alone, and that cancer would beresponsible for nearly 600,000 U.S. deaths.

Cancer is characterized by the unregulated proliferation of cells, whichdisrupt the function of tissues. The proliferation of cells can becaused by an abnormal increase in cell production or a disruption in thecell death pathway. In any event, disruptors of cell function can impactthe proliferation of cells and in particular cancer cells by reducing orinhibiting cell proliferation. For example, the modulation of cancercell metabolism can lead to the reduction or inhibition of cellproliferation. Equally, compounds of the invention may reduce, disrupt,or inhibit the growth or proliferation of a cancer cell or it may inducethe death of a cancer cell. As such, cancer cell metabolism, andreducing cell proliferation, is a potential target for disrupting cancergrowth and ultimately a therapeutic pathway for cancer treatment.Accordingly, the certain embodiments of the invention contemplatecompounds that modulate cancer cell metabolism and/or reduce cellproliferation. Reduction in cell proliferation could be achieved eitherby increasing cell death or by reducing the rate of cell growth.

It has been observed that certain compounds having antibiotic activityhave a beneficial effect when administered to patients with cancer. Theinventors have found that compounds having a phosphonium ion linked toazithromycin are able to modulate cancer cell metabolism in cancer celllines and, accordingly, prevent and/or treat cancer.

The “prevention” of cancer may be taken as including the prevention ofthe formation of tumours, including primary tumours, metastatic tumours,or tumours associated with cancer onset, resistance or relapse. Theprevention of cancer may also be taken as encompassing the prevention ofthe progression of cancer. In this context, prevention of development ofcancer may be demonstrated by preventing an increase in the “stage” of atumour (using an appropriate cancer staging method) that has beentreated using the compounds of the invention. The prevention of increasein cancer stage may be compared to progression of an untreated tumour,or compared to the extent of progression that would be expected by aclinician in the event that the tumour was not treated.

The “treatment” of cancer may be taken as including any improvement ofpathology, symptoms or prognosis that is achieved in respect of cancerin a subject receiving compounds of the invention. Treatment may beindicated by a partial improvement of such indications, or by a totalimprovement (e.g. the absence of cancer following medical use of thecompounds of the invention).

Accordingly, the prevention and/or treatment as defined above areintended aims of certain embodiments of the invention. The abovedefinitions of treatment or prevention of cancer apply equally to thespecific forms of cancer that are also contemplated.

Recent developments in cancer therapy have suggested that certainantibiotic compounds may be useful in cancer treatment. The mechanismsby which these agents, which include the antibiotics azithromycin anddoxycycline, exert a therapeutic effect have been open to markedlydifferent explanations. Some authors have suggested that these agentsinhibit matrix metalloproteinases (MMPs) and thereby achieve ananti-inflammatory effect, while others suggest that they impair thecells' response to DNA damage, thereby increasing the effectiveness ofchemotherapy or radiotherapy on bulk tumour cells. Still other articleshave indicated that the antibiotics target mitochondrial function.

However, there have also been reports that antibiotic use can increaserisk of colorectal cancer. A recent study identified that increasingduration of antibiotic use was significantly associated with anincreased risk of colorectal adenoma (Cao Y, Wu K, Mehta R, et al,“Long-term use of antibiotics and risk of colorectal adenoma”, Gut,2017, 0, page 1-7).

Surprisingly, the inventors have found that the compounds having aphosphonium ion linked to azithromycin have increased activity againstcertain cancer cell lines compared to azithromycin itself. This isdemonstrated by the reduced cancer cell proliferation observed in a cellconfluence assay.

Also provided is a method of preventing and/or treating cancer in asubject needing such prevention and/or treatment, the method comprisingadministering a therapeutically effective amount of a compound of theinvention to the subject. A therapeutically effective amount of acompound of the invention may be an amount of such a compound sufficientto treat a variety of cancers, including the modulation of cancer cellsor other dysfunctional cells (such as tumour initiating cells, stem-likecancer cells, cancer stem cells, or a population of cells with stemcell-like features that exist in tumors and that give rise to the bulkof tumor cells with more differentiated phenotypes). References tocancer cells include hybrid and giant cells. It will be appreciated thatthe therapeutically effective amount of the compound of the inventionmay be provided in a single incidence of administration, or cumulativelythrough multiple incidences of administration.

The same considerations regarding the types of cancers to be treated,and benefits provided by treatment, described with respect to themedical uses of the compounds of the invention also apply to the methodsof treatment of the invention.

BRIEF SUMMARY OF THE DISCLOSURE

In a first aspect of the invention there is provided a compoundcomprising an ion of formula (I) or a pharmaceutically acceptable saltthereof:

whereineither Z¹ is

and Z² is R^(4b);

or Z² is

and Z¹ is R^(2b);Z³ is independently selected from H, C(O)—C₁-C₆-alkyl or Z³ has thestructure:

Z⁴ is independently selected from H, C(O)—C₁-C₆-alkyl or Z⁴ has thestructure:

-L¹- is independently absent or is selected from —C(O)—, —C(O)O—,—S(O)₂—, —S(O)—, —C(O)NR⁵, and —S(O)₂NR⁵—;-L³- is independently at each occurrence either absent or selected from:—O—, —S—, —NR⁶—, —C(O)—, —OC(O)—, —C(O)O—, —S(O)₂—, —S(O)—, —NR⁵C(O)—,—C(O)NR⁵, —NR⁵S(O)₂—, —S(O)₂NR⁵—, —OC(O)NR⁵—, —NR⁵C(O)O—, NR⁵C(O)NR⁵,—CR⁷═CR⁷— and —C≡C—;-L²- and -L⁴- are each independently at each occurrence—C₁-C₄-alkylene-, each alkylene group being unsubstituted or substitutedwith from 1 to 6 independently selected R⁸ groups; provided that any-L₂- or-L₄- group that is attached at each end to an atom selected fromoxygen, nitrogen, sulphur or phosphorous is —C₂-C₄-alkylene-;-L⁵- is independently absent or is selected from —C(O)— and —C(O)NR⁵—;n is an integer selected from 0, 1, 2, 3, 4 and 5;wherein L¹, L², L³, L⁴, L⁵ and n are selected such that length of thelinker formed by those groups is from 3 to 16 atoms;R^(1a), R^(1b) and R^(1c) are each unsubstituted phenyl;R^(2a), R^(2b) and R^(2c) are each independently selected from H andC₁-C₆-alkyl;R^(3a) is independently selected from: H, C₁-C₆-alkyl andC(O)—C₁-C₆-alkyl;R^(3b) and R^(3c) are each independently selected from: H andC(O)—C₁-C₆-alkyl;R^(4a) and R^(4b) are each independently selected from: H, C₁-C₆-alkyland C(O)—C₁-C₆-alkyl; orR^(4a) and R^(4b) taken together form C(O);R⁵ is independently at each occurrence selected from H and C₁-C₆-alkyl;R⁶ and R¹¹ are each independently at each occurrence selected from: H,C₁-C₆-alkyl, C(O)C₁-C₆-alkyl and S(O)₂—C₁-C₆-alkyl;R⁷ is independently at each occurrence selected from H, C₁-C₄-alkyl andhalo;R⁸ is independently at each occurrence selected from: C₁-C₆-alkyl,C₂-C₆-alkynyl, C₂-C₆-alkenyl, C₁-C₆-haloalkyl, OR⁹, SR¹⁰, NR¹⁰R¹¹,C(O)OR¹⁰, C(O)NR¹⁰R¹⁰, NR¹⁰C(O)OR¹⁰, OR¹⁰C(O)NR¹⁰R¹⁰, halo, cyano,nitro, C(O)R¹⁰, S(O)₂OR¹⁰, S(O)₂R¹⁰, S(O)R¹⁰ and S(O)₂NR¹⁰R¹⁰;R⁹ is independently at each occurrence selected from: H, C₁-C₆-alkyl andC₁-C₆-haloalkyl;R¹⁰ is independently at each occurrence selected from: H andC₁-C₆-alkyl;and wherein any of the abovementioned alkyl, alkenyl, alkynyl,cycloalkyl, heterocycloalkyl, heteroaryl or phenyl groups is optionallysubstituted where chemically allowable by from 1 to 4 groupsindependently selected from oxo, C₁-C₆-alkyl, C₂-C₆-alkynyl,C₂-C₆-alkenyl, C₁-C₆-haloalkyl, OR^(a), NR^(a)R^(b), SR^(a), C(O)OR^(a),C(O)NR^(a)R^(a), halo, cyano, nitro, C(O)R^(a), S(O)₂OR^(a), S(O)₂R^(a),S(O)R^(a) and S(O)₂NR^(a)R^(a); wherein R^(a) is independently at eachoccurrence selected from: H and C₁-C₆-alkyl; and R^(b) is independentlyat each occurrence selected from: H, C₁-C₆-alkyl, C(O)C₁-C₆-alkyl andS(O)₂—C₁-C₆-alkyl.

For the absence of doubt, where n is greater than 1, each -L³-L⁴- unitis selected independently of the other each -L³-L⁴- unit or -L³-L⁴-units. Thus, each -L³-L⁴- unit may be the same or they may be different.

For the absence of doubt the atom length of the linkers formed by L¹,L², L³ L⁴ and L⁵ is the number of atoms in a straight chain from thephosphorous atom of the phosphonium to the oxygen or nitrogen atom viawhich the linker is attached to the azithromycin portion of the ions.The length does not include any substituents or branching that might bepresent on the chain.

For the absence of doubt where a bivalent group (e.g. L¹, L², L³, L⁴, L⁵or a combination thereof) is represented in text, the left hand portionof the linker group is attached, either directly or indirectly, to theoxygen or nitrogen atom via which the linker is attached to theazithromycin portion of the ions and the right hand portion of thelinker group is attached, either directly or indirectly, to thephosphorous atom of the phosphonium.

A compound comprising an ion of formula (I) or a pharmaceuticallyacceptable salt thereof:

whereineither Z¹ is R

and Z² is R^(4b);

or Z² is

and Z¹ is R^(2b);Z³ is independently selected from H, C(O)—C₁-C₆-alkyl or Z³ has thestructure:

Z⁴ is independently selected from H, C(O)—C₁-C₆-alkyl or Z⁴ has thestructure:

-L¹- is independently absent or is selected from —C(O)—, —C(O)O—,—S(O)₂—, —S(O)—, —C(O)NR⁵, and —S(O)₂NR⁵—;-L³- is independently at each occurrence either absent or selected from:—O—, —S—, —NR⁶—, —C(O)—, —OC(O)—, —C(O)O—, —S(O)₂—, —S(O)—, —NR⁵C(O)—,—C(O)NR⁵, —NR⁵S(O)₂—, —S(O)₂NR⁵—, —OC(O)NR⁵—, —NR⁵C(O)O—, NR⁵C(O)NR⁵,—CR⁷═CR⁷— and —C≡C—;-L²- and -L⁴- are each independently at each occurrence—C₁-C₄-alkylene-, each alkylene group being unsubstituted or substitutedwith from 1 to 6 independently selected R⁸ groups; provided that any-L₂- or -L₄- group that is attached at each end to an atom selected fromoxygen, nitrogen, sulphur or phosphorous is —C₂-C₄-alkylene-;-L⁵- is independently absent or is selected from —C(O)— and —C(O)NR⁵—;n is an integer selected from 0, 1, 2, 3, 4 and 5;wherein L¹, L², L³, L⁴, L⁵ and n are selected such that length of thelinker formed by those groups is from 3 to 16 atoms;R^(1a), R^(1b) and R^(1c) are each unsubstituted phenyl;R^(2a), R^(2b) and R^(2c) are each independently selected from H andC₁-C₆-alkyl;R^(3a) is independently selected from: H, C₁-C₆-alkyl andC(O)—C₁-C₆-alkyl;R^(3b) and R^(3c) are each independently selected from: H andC(O)—C₁-C₆-alkyl;R^(4a) and R^(4b) are each independently selected from: H, C₁-C₆-alkyland C(O)—C₁-C₆-alkyl; orR^(4a) and R^(4b) taken together form C(O);R⁵ is independently at each occurrence selected from H and C₁-C₆-alkyl;R⁶ and R¹¹ are each independently at each occurrence selected from: H,C₁-C₆-alkyl, C(O)C₁-C₆-alkyl and S(O)₂—C₁-C₆-alkyl;R⁷ is independently at each occurrence selected from H, C₁-C₄-alkyl andhalo;R⁸ is independently at each occurrence selected from: C₁-C₆-alkyl,C₂-C₆-alkynyl, C₂-C₆-alkenyl, C₁-C₆-haloalkyl, OR⁹, SR¹⁰, NR¹⁰R¹¹,C(O)OR¹⁰, C(O)NR¹⁰R¹⁰, NR¹⁰C(O)OR¹⁰, OR¹⁰C(O)NR¹⁰R¹⁰, halo, cyano,nitro, C(O)R¹⁰, S(O)₂OR¹⁰, S(O)₂R¹⁰, S(O)R¹⁰ and S(O)₂NR¹⁰R¹⁰;R⁹ is independently at each occurrence selected from: H, C₁-C₆-alkyl andC₁-C₆-haloalkyl;R¹⁰ is independently at each occurrence selected from: H andC₁-C₆-alkyl;and wherein any of the abovementioned alkyl, alkenyl, alkynyl,cycloalkyl, heterocycloalkyl, heteroaryl or phenyl groups is optionallysubstituted where chemically allowable by from 1 to 4 groupsindependently selected from oxo, C₁-C₆-alkyl, C₂-C₆-alkynyl,C₂-C₆-alkenyl, C₁-C₆-haloalkyl, OR^(a), NR^(a)R^(b), SR^(a), C(O)OR^(a),C(O)NR^(a)R^(a), halo, cyano, nitro, C(O)R^(a), S(O)₂OR^(a), S(O)₂R^(a),S(O)R^(a) and S(O)₂NR^(a)R^(a); wherein R^(a) is independently at eachoccurrence selected from: H and C₁-C₆-alkyl; and R^(b) is independentlyat each occurrence selected from: H, C₁-C₆-alkyl, C(O)C₁-C₆-alkyl andS(O)₂—C₁-C₆-alkyl.

In embodiments the ion of formula (I) is an ion of formula (II):

In embodiments the ion of formula (I) is an ion of formula (IIIa) and(IIIb):

In embodiments, the ion of formula (I) is an ion of formula (IVa) and(IVb):

In embodiments the ion of formula (I) is an ion of formula (Va) and(Vb):

wherein L⁶ is a C₂-C₁₅-alkylene group optionally substituted with from 0to 10 R⁸ groups.

In embodiments, the ion of formula (I) is an ion of formula (VIa) and(VIb):

wherein L⁷ is a C₃-C₁₆-alkylene group optionally substituted with from 0to 10 R⁸ groups.

In embodiments the ion of formula (I) is an ion of formula (VIIa) and(VIIb):

wherein L⁸ is a C₂-C₁₃-alkylene group optionally substituted with from 0to 10 R⁸ groups.

The following statements apply to compounds of any of formulae (I) to(VIIb). These statements are independent and interchangeable. In otherwords, any of the features described in any one of the followingstatements may (where chemically allowable) be combined with thefeatures described in one or more other statements below. In particular,where a compound is exemplified or illustrated in this specification,any two or more of the statements below which describe a feature of thatcompound, expressed at any level of generality, may be combined so as torepresent subject matter which is contemplated as forming part of thedisclosure of this invention in this specification.

In embodiments, Z¹ is

and Z² is R^(4b).

In embodiments, Z² is

and Z¹ is R^(2b).

In embodiments, Z³ is

In embodiments, Z³ is

In embodiments, Z³ is

In embodiments, Z³ is H.

In embodiments, Z³ is

wherein R^(3c) is H.

In embodiments, Z³ is

wherein R^(3c) is H.

In embodiments, Z⁴ is

or H.

In embodiments, Z⁴ is Me.

In embodiments, Z⁴ is H.

In embodiments, R^(2a) is C₁-C₆-alkyl.

In embodiments, R^(2a) is methyl.

In embodiments, R^(2c) is C₁-C₆-alkyl.

In embodiments, R^(2c) is methyl.

In embodiments, R^(2a) is C₁-C₆-alkyl and R^(2c) is C₁-C₆-alkyl.

In embodiments, R^(2a) is methyl and R^(2c) is methyl.

In embodiments, R^(2b) (when present) is methyl.

In embodiments, R^(2a) is methyl, R^(2b) (when present) is methyl andR^(2c) is methyl.

In embodiments, R^(3a) is H.

In embodiments, R^(3b) is H or C(O)CH₃.

In embodiments, R^(3b) is H.

In embodiments, R^(3b) is C(O)CH₃.

In embodiments, R^(3c) is H.

In embodiments, R^(3a) is H and R^(3b) is H.

In embodiments, R^(3a) is H and R^(3b) is C(O)CH₃.

In embodiments, R^(3a) is H, R^(3b) is C(O)CH₃ and R^(3c) is H.

In embodiments, R^(3a) is H, R^(3b) is H and R^(3c) is H.

In embodiments, R^(4a) is H.

In embodiments, R^(4b) is H.

In embodiments, R^(4a) is H and R^(4b) is H.

In embodiments, R^(4a) and R^(4b) together form C(O).

In embodiments, Z³ is

R^(3c) is H, R^(2a) is C₁-C₆-alkyl; R^(2b) is C₁-C₆-alkyl; R^(3a) is Hand R^(3b) is H.

In embodiments, Z³ is

R^(3c) is H, R^(2a) is methyl; R^(2c) is methyl; R^(3a) is H and R^(3b)is H.

In embodiments, Z³ is

R^(3c) is H, R^(2a) is C₁-C₆-alkyl; R^(2c) is C₁-C₆-alkyl; R^(3a) is H;R^(3b) is H; R^(4a) is H and R^(4b) (when present) is H.

In embodiments, Z³ is

R^(3c) is H, R^(2a) is methyl; R^(2c) is methyl; R^(3a) is H; R^(3b) isH, R^(4a) is H and R^(4b) (when present) is H.

In embodiments, Z⁴ is

R^(2a) is C₁-C₆-alkyl; R^(2b) is C₁-C₆-alkyl; R^(3a) is H and R^(3b) isH.

In embodiments, Z⁴ is

R^(2a) is methyl; R^(2c) is methyl; R^(3a) is H and R^(3b) is H.

In embodiments, Z⁴ is

R^(2a) is C₁-C₆-alkyl; R^(2c) is C₁-C₆-alkyl; R^(3a) is H; R^(3b) is H;R^(4a) is H and R^(4b) (when present) is H.

In embodiments, Z⁴ is

R^(2a) is methyl; R^(2c) is methyl; R^(3a) is H; R^(3b) is H, R^(4a) isH and R^(4b) (when present) is H.

In embodiments, Z³ is H and Z⁴ is H.

In embodiments, Z³ is

and Z⁴ is Me, optionally wherein R^(3c) is H, R^(2a) is C₁-C₆-alkyl;R^(2c) is C₁-C₆-alkyl; R^(3a) is H and R^(3b) is H.

In embodiments, Z³ is H, R^(2a) is C₁-C₆-alkyl; R^(2b) is C₁-C₆-alkyl;R^(3a) is H and R^(3b) is H.

In embodiments, Z³ is H, R^(2a) is methyl; R^(2c) is methyl; R^(3a) is Hand R^(3b) is H.

In embodiments, Z³ is H, R^(2a) is C₁-C₆-alkyl; R^(2c) is C₁-C₆-alkyl;R^(3a) is H; R^(3b) is H; R^(4a) is H and R^(4b) (when present) is H.

In embodiments, Z³ is H, R^(2a) is methyl; R^(2c) is methyl; R^(3a) isH; R^(3b) is H, R^(4a) is H and R^(4b) (when present) is H.

In embodiments, Z⁴ is H, R^(2a) is C₁-C₆-alkyl; R^(2b) is C₁-C₆-alkyl;R^(3a) is H and R^(3b) is H.

In embodiments, Z⁴ is H, R^(2a) is methyl; R^(2c) is methyl; R^(3a) is Hand R^(3b) is H.

In embodiments, Z⁴ is H, R^(2a) is C₁-C₆-alkyl; R^(2c) is C₁-C₆-alkyl;R^(3a) is H; R^(3b) is H; R^(4a) is H and R^(4b) (when present) is H.

In embodiments, Z⁴ is H, R^(2a) is methyl; R^(2c) is methyl; R^(3a) isH; R^(3b) is H, R^(4a) is H and R^(4b) (when present) is H.

In embodiments, R⁵ is at any particular occurrence H. In embodiments, R⁵is at each occurrence H.

In embodiments, R⁵ is at any particular occurrence C₁-C₄-alkyl, e.g.methyl. In embodiments, R⁵ is at each occurrence C₁-C₄-alkyl, e.g.methyl.

In embodiments, R⁶ is at any particular occurrence H. In embodiments, R⁶is at each occurrence H.

In embodiments, R⁶ is at any particular occurrence C₁-C₄-alkyl, e.g.methyl. In embodiments, R⁶ is at each occurrence C₁-C₄-alkyl, e.g.methyl.

In embodiments, R⁷ is at any particular occurrence H. In embodiments, R⁷is at each occurrence H.

In embodiments, R⁸ is at any particular occurrence C₁-C₄-alkyl, e.g.methyl. In embodiments, R⁸ is at each occurrence C₁-C₄-alkyl, e.g.methyl.

In embodiments, R⁹ is at any particular occurrence H. In embodiments, R⁹is at each occurrence H.

In embodiments, R⁹ is at any particular occurrence C₁-C₄-alkyl, e.g.methyl. In embodiments, R⁹ is at each occurrence C₁-C₄-alkyl, e.g.methyl.

In embodiments, R¹⁰ is at any particular occurrence H. In embodiments,R¹⁰ is at each occurrence H.

In embodiments, R¹⁰ is at any particular occurrence C₁-C₄-alkyl, e.g.methyl. In embodiments, R¹⁰ is at each occurrence C₁-C₄-alkyl, e.g.methyl.

In embodiments, R¹¹ is at any particular occurrence H. In embodiments,R¹¹ is at each occurrence H.

In embodiments, R¹¹ is at any particular occurrence C₁-C₄-alkyl, e.g.methyl. In embodiments, R¹⁰ is at each occurrence C₁-C₄-alkyl, e.g.methyl.

In embodiments, L¹, L², L³, L⁴, L⁵ and n are selected such that lengthof the linker formed by those groups is from 8 to 14 atoms.

In embodiments. Z¹ is

and Z² is R^(4b).

In embodiments, L¹ is selected from —C(O)— and —S(O)₂—. In embodiments,L¹ is —C(O)—.

In embodiments, L¹ is absent.

In certain embodiments, L³ is at each occurrence absent. Thus, the group-L²-(L³-L⁴)_(n)- may form an alkylene linker group.

In certain embodiments, L³ is at each occurrence —O— and -L⁴- is at eachoccurrence —C₂-C₄-alkylene-. Thus, the group -(L³-L⁴)_(n)- may form aether or polyether linker group. -L⁴- may at each occurrence represent—CH₂CH₂— or —CH₂CH₂CH₂—. Thus, the group -(L³-L⁴)_(n)- may form a,ethylene glycol, polyethyleneglycol, propyleneglycol or polypropyleneglycol linker group.

In certain embodiments, L³ is at each occurrence —NR⁵C(O)—, —C(O)NR⁵.Thus, the group -(L³-L⁴)_(n)- may form a peptide linker group. In theseembodiments, it may be that -L⁴- is at each occurrence —C₁-alkylene-.

In embodiments, L¹, L², L³, L⁴ and n are selected such that length ofthe linker formed by those groups is from 8 to 14 atoms.

In embodiments Z¹ is:

wherein L⁶ is a C₂-C₁₅-alkylene group optionally substituted with from 0to 10 R⁸ groups. L⁶ may be a C₇-C₁₃-alkylene group optionallysubstituted with from 0 to 10 R⁸ groups. L⁶ may be unsubstituted.

In specific embodiments, -L¹-L²-(L³-L⁴-)_(n) represents —C(O)(CH₂)₁₁—,—C(O)(CH₂)₁₀—, —C(O)(CH₂)₉—, —C(O)(CH₂)₈—, —C(O)(CH₂)₇—, —C(O)(CH₂)₆—,or —C(O)(CH₂)₅—.

In specific embodiments, -L¹-L²-(L³-L⁴)_(n)- represents —C(O)NH(CH₂)₈—,—C(O)NH(CH₂)₇—, or —C(O)NH(CH₂)₆—.

In embodiments Z¹ is:

wherein L⁷ is a C₃-C₁₆-alkylene group optionally substituted with from 0to 10 R⁸ groups. L⁷ may be a C₈-C₁₄-alkylene group optionallysubstituted with from 0 to 10 R⁸ groups. L⁷ may be unsubstituted.

In embodiments L⁵ is absent.

In embodiments, L⁵ is —C(O)—.

In embodiments, L⁵ is-C(O)NR⁵—, e.g. —C(O)NH—.

In certain embodiments, L³ is at each occurrence absent. Thus, the group-L²-(L³-L⁴)_(n)- may form an alkylene linker group.

In certain embodiments, L³ is at each occurrence —O— and -L⁴- is at eachoccurrence —C₂-C₄-alkylene-. Thus, the group -(L³-L⁴)_(n)- may form aether or polyether linker group. -L⁴- may at each occurrence represent—CH₂CH₂— or —CH₂CH₂CH₂—. Thus, the group -(L³-L⁴)_(n)- may form a,ethylene glycol, polyethyleneglycol, propyleneglycol or polypropyleneglycol linker group.

In certain embodiments, L³ is at each occurrence —NR⁵C(O)—, —C(O)NR⁵.Thus, the group -(L³-L⁴)_(n)- may form a peptide linker group. In theseembodiments, it may be that -L⁴- is at each occurrence —C₁-alkylene-.

In embodiments, L⁵, L², L³, L⁴ and n are selected such that length ofthe linker formed by those groups is from 8 to 14 atoms.

In embodiments Z² is:

wherein L⁸ is a C₂-C₁₃-alkylene group optionally substituted with from 0to 10 R⁸ groups. L⁸ may be a C₅-C₁₁-alkylene group optionallysubstituted with from 0 to 10 R⁸ groups. L⁸ may be unsubstituted.

In embodiments of Z², -L⁵-L²-(L³-L⁴)_(n)- represents —C(O)NH(CH₂)₁₁—,—C(O)NH(CH₂)₁₀—, —C(O)NH(CH₂)₉—, —C(O)NH(CH₂)₈—, —C(O)NH(CH₂)₇—,—C(O)NH(CH₂)₆—, or —C(O)NH(CH₂)₅—.

In embodiments of Z², -L⁵-L²-(L³-L⁴)_(n)- represents —C(O)NH(CH₂)₁₀—,—C(O)NH(CH₂)₈—, or —C(O)NH(CH₂)₆—.

In embodiments n is an integer selected from 0 or 1.

In an embodiment of the invention the ion of formula (I) is a ionselected from:

The cation of formula (I) will be associated with an anionic counterion. For administration to a subject, the cation of formula (I) will beassociated with a pharmaceutically acceptable anionic counterion. Thefirst aspect of the invention also, therefore, provides a compoundcomprising the ion of formula (I) and a pharmaceutically acceptableanion. The anion may have a single negative charge. For example theanion may be selected from: halo (e.g. Cl, Br and I), BF₄, PF₆,CF₃C(O)O, HC(O)O, HCO₃, (CF₃SO₂)₂N, (C₂F₅)₃PF₃, HSO₄, C₁-C₁₅-alkylSO₄,CH₃C(O)O, CF₃SO₃, Tosyl-O, C(CN)₃, N(CN)₂ or the carboxylate anion of aproteinogenic amino acid. For the avoidance of doubt each anion listedin the preceding sentence possesses a single negative charge. The anionmay have multiple negative charges, e.g. PO₄ ³⁻ or CO₃ ²⁻. The anion maybe derived from a di- or tri-acid, e.g. glutamic acid, succinic acid,malic acid, citric acid, tartaric acid. It may be a mono-carboxylate ofsaid di- or tri-acid. The remaining carboxylic acid groups may be in theform of protonated carboxylic acids, C₁-C₁₂-alkylesters, or they maylikewise be carboxylate anions. Said carboxylate anions may each beaccompanied by a pharmaceutically acceptable metal cation or by anothercation of formula (I).

The anions associated with the cations of the invention can be quitelabile. It may be therefore that the cation of the invention is presentassociated with two or more different anions. Ion exchange processes canbe used to control the identity of the anion associated with the cationof the invention.

In embodiments the anion is Cl, Br, I, PF₆, CF₃C(O)O, or HC(O)O.

In an aspect of the invention, the compounds of the invention are formedical use.

In an aspect the compounds of the first aspect of the invention are foruse in the treatment of cancer. The compounds may also be for use inreducing cell proliferation of abnormal cells, such as cancer cells.

In an embodiment the compounds of the first aspect of the invention arefor use in the treatment of solid tumours and other cancers, e.g.cancers classed as not being solid cancers. Amongst cancers that can betreated by the compounds of the invention are: leukaemia, lymphoma,sarcoma, or carcinoma.

In a further aspect of the invention there is provided a method for thetreatment of cancer, wherein the method comprises the administration ofa therapeutically effective amount of a compound of the first aspect ofthe invention. The method may also be for use in reducing cellproliferation of abnormal cells, such as cancer cells.

In an embodiment the method is for the treatment of solid tumours andother cancers, e.g. cancers classed as not being solid cancers. Amongstcancers that can be treated by the methods of the invention are:leukaemia, lymphoma, sarcoma, or carcinoma.

The “treatment” of cancer may be taken to include prevention. Treatmentalso encompasses including any improvement of pathology, symptoms orprognosis that is achieved in respect of cancer in a subject receivingcompounds of the invention. Treatment may be indicated by a partialimprovement of such indications, or by a total improvement (e.g. theabsence of cancer following medical use of the compounds of theinvention).

The “prevention” of cancer may be taken as including the prevention ofthe formation of new tumours, including new primary tumours or newmetastatic tumours. The prevention of cancer may also be taken asencompassing the prevention of the progression of cancer. In thiscontext, prevention of development of cancer may be demonstrated bypreventing an increase in the “stage” of a tumour (using an appropriatecancer staging method) that has been treated using the compounds of theinvention. The prevention of increase in cancer stage may be compared toprogression of an untreated tumour, or compared to the extent ofprogression that would be expected by a clinician in the event that thetumour was not treated.

The compounds of the first aspect of the invention may be for use inincreasing cancer cell death or for decreasing cell proliferation byanother mechanism, such as inhibiting cell replication. The compoundsmay be used for this purpose in vitro or in vivo.

The compounds of the invention may be for use in the modulation ofcancer cells or other dysfunctional cells (such as tumour initiatingcells, stem-like cancer cells, cancer stem cells, or a population ofcells with stem cell-like features that exist in tumors and that giverise to the bulk of tumor cells with more differentiated phenotypes).Accordingly, there is provided a method of modulating cancer cells orother dysfunctional cells in vivo or in vitro by exposing the cancercells or other dysfunctional cells to a compound of the first aspect ofthe invention. The compound may be exposed to the cancer cells or otherdysfunctional cells in an effective amount, for example atherapeutically effective amount such as in the case of a method oftreatment or an in vivo method.

In another aspect of the invention there is provided a pharmaceuticalcomposition, wherein the composition comprises a compound of theinvention and one or more pharmaceutically acceptable excipients.

In an embodiment the pharmaceutical composition may be a combinationproduct comprising one or more different pharmaceutically active agents.The one or more additional pharmaceutically active agents may be ananti-cancer agent described below. The one or more pharmaceuticallyactive agents may independently be selected from a different therapeuticclass, e.g. antibiotic, anti-viral, anti-emetic, pain management, etc.

The present invention also contemplates the subject matter contained inthe following numbered clauses:

1. A compound comprising an ion of formula (I) or a pharmaceuticallyacceptable salt thereof:

whereineither Z¹ is

and Z² is R^(4b);

or Z² is

and Z¹ is R^(2b);Z³ is independently selected from H, C(O)—C₁-C₆-alkyl or Z³ has thestructure:

Z⁴ is independently selected from H, C(O)—C₁-C₆-alkyl or Z⁴ has thestructure:

-L¹- is independently absent or is selected from —C(O)—, —C(O)O—,—S(O)₂—, —S(O)—, —C(O)NR⁵, and —S(O)₂NR⁵—;-L³- is independently at each occurrence either absent or selected from:—O—, —S—, —NR⁶—, —C(O)—, —OC(O)—, —C(O)O—, —S(O)₂—, —S(O)—, —NR⁵C(O)—,—C(O)NR⁵, —NR⁵S(O)₂—, —S(O)₂NR⁵—, —OC(O)NR⁵—, —NR⁵C(O)O—, NR⁵C(O)NR⁵,—CR⁷═CR⁷— and —C≡C—;-L²- and -L⁴- are each independently at each occurrence—C₁-C₄-alkylene-, each alkylene group being unsubstituted or substitutedwith from 1 to 6 independently selected R⁸ groups; provided that any-L₂- or -L₄- group that is attached at each end to an atom selected fromoxygen, nitrogen, sulphur or phosphorous is —C₂-C₄-alkylene-;-L⁵- is independently absent or is selected from —C(O)— and —C(O)NR⁵—;n is an integer selected from 0, 1, 2, 3, 4 and 5;wherein L¹, L², L³, L⁴, L⁵ and n are selected such that length of thelinker formed by those groups is from 3 to 16 atoms;R^(1a), R^(1b) and R^(1c) are each unsubstituted phenyl;R^(2a), R^(2b) and R^(2c) are each independently selected from H andC₁-C₆-alkyl;R^(3a) is independently selected from: H, C₁-C₆-alkyl andC(O)—C₁-C₆-alkyl;R^(3b) and R^(3c) are each independently selected from: H andC(O)—C₁-C₆-alkyl;R^(4a) and R^(4b) are each independently selected from: H, C₁-C₆-alkyland C(O)—C₁-C₆-alkyl; orR^(4a) and R^(4b) taken together form C(O);R⁵ is independently at each occurrence selected from H and C₁-C₆-alkyl;R⁶ and R¹¹ are each independently at each occurrence selected from: H,C₁-C₆-alkyl, C(O)C₁-C₆-alkyl and S(O)₂—C₁-C₆-alkyl;R⁷ is independently at each occurrence selected from H, C₁-C₄-alkyl andhalo;R⁸ is independently at each occurrence selected from: C₁-C₆-alkyl,C₂-C₆-alkynyl, C₂-C₆-alkenyl, C₁-C₆-haloalkyl, OR⁹, SR¹⁰, NR¹⁰R¹¹,C(O)OR¹⁰, C(O)NR¹⁰R¹⁰, NR¹⁰C(O)OR¹⁰, OR¹⁰C(O)NR¹⁰R¹⁰, halo, cyano,nitro, C(O)R¹⁰, S(O)₂OR¹⁰, S(O)₂R¹⁰, S(O)R¹⁰ and S(O)₂NR¹⁰R¹⁰;R⁹ is independently at each occurrence selected from: H, C₁-C₆-alkyl andC₁-C₆-haloalkyl;R¹⁰ is independently at each occurrence selected from: H andC₁-C₆-alkyl;and wherein any of the abovementioned alkyl, alkenyl, alkynyl,cycloalkyl, heterocycloalkyl, heteroaryl or phenyl groups is optionallysubstituted where chemically allowable by from 1 to 4 groupsindependently selected from oxo, C₁-C₆-alkyl, C₂-C₆-alkynyl,C₂-C₆-alkenyl, C₁-C₆-haloalkyl, OR^(a), NR^(a)R^(b), SR^(a), C(O)OR^(a),C(O)NR^(a)R^(a), halo, cyano, nitro, C(O)R^(a), S(O)₂OR^(a), S(O)₂R^(a),S(O)R^(a) and S(O)₂NR^(a)R^(a); wherein R^(a) is independently at eachoccurrence selected from: H and C₁-C₆-alkyl; and R^(b) is independentlyat each occurrence selected from: H, C₁-C₆-alkyl, C(O)C₁-C₆-alkyl andS(O)₂—C₁-C₆-alkyl.2. A compound of clause 1, wherein Z¹ is

R^(a) and Z² is R^(4b).3. A compound of clause 2, wherein L¹ is selected from —C(O)— and—S(O)₂—.4. A compound of clause 2, wherein L¹ is absent.5. A compound of any one of clauses 2 to 4, wherein R^(4a) is H andR^(4b) is H.6. A compound of any one of clauses 2 to 4, wherein R^(4a) and R^(4b)together form C(O).7. A compound of clause 1, wherein Z² is

and Z¹ is R^(2b), preferably wherein R^(2b) is methyl.8. A compound of clause 7, wherein L⁵ is —C(O)—.9. A compound of clause 7, wherein L⁵ is —C(O)NR⁵—.10. A compound of any one of clauses 7 to 9, wherein R^(4a) is H.11. A compound of any one of clauses 1 to 10, wherein L³ is at eachoccurrence absent.12. A compound of any one of clauses 1 to 11, wherein L¹, L², L³, L⁴, L⁵and n are selected such that length of the linker formed by those groupsis from 8 to 14 atoms.13. A compound of any one of clauses 1 to 12, wherein Z³ is

preferably wherein R^(3c) is H.14. A compound of any one of clauses 1 to 13, wherein R^(2a) is methyland R^(2c) is methyl.15. A compound of any one of clauses 1 to 14, wherein R^(3a) is H andR^(3b) is H.16. A compound of any one of clauses 1 to 14, wherein R^(3a) is H andR^(3b) is C(O)CH₃.17. A compound of any one of clauses 1 to 16, wherein the compound isfor medical use.18. A compound of any one of clauses 1 to 16, wherein the compound isfor use in the treatment of cancer.19. A method for the treatment of cancer, wherein the method comprisesthe administration of a therapeutically effective amount of a compoundof any one of clauses 1 to 16.20. A pharmaceutical composition, wherein the composition comprises acompound of any one of clauses 1 to 16 and one or more pharmaceuticallyacceptable excipients.

DETAILED DESCRIPTION

Given below are definitions of terms used in this application. Any termnot defined herein takes the normal meaning as the skilled person wouldunderstand the term.

The term “halo” or “halogen” refers to an atom selected from fluorine,chlorine, bromine and iodine. “Halo” or “halogen” may refer to an atomselected from Cl and F. “Halo” or “halogen” may refer to fluorine.

The term “alkyl” refers to a linear or branched hydrocarbon chain. Theterm “C₁-C₈ alkyl” refers to a linear or branched hydrocarbon chaincontaining 1, 2, 3, 4, 5, 6, 7 or 8 carbon atoms. The term “C₁-C₆ alkyl”refers to a linear or branched hydrocarbon chain containing 1, 2, 3, 4,5 or 6 carbon atoms. The term “C₁-C₆ alkyl” for example refers tomethyl, ethyl, n-propyl, iso-propyl, n-butyl, sec-butyl, tert-butyl,n-pentyl and n-hexyl. The “alkyl” group may be substituted orunsubstituted by one or more substituents. Substituents for the alkylgroup may be halo (for example fluorine, chlorine, bromine and iodine),OH and C₁-C₆ alkoxy. In addition, alkylene groups may be linear orbranched and may have two places of attachment to the remainder of themolecule.

The term “alkylene” refers to a divalent group which is a linear orbranched hydrocarbon chain. With the “alkylene” group being divalent,the group must form two bonds to other groups. The term “C₁-C₈-alkylene”may refer to —CH₂—, —CH₂CH₂—, —CH₂CH₂CH₂—, —CH₂CH₂CH₂CH₂—,—CH₂CH₂CH₂CH₂CH₂—, —CH₂CH₂CH₂CH₂CH₂CH₂—, —CH₂CH₂CH₂CH₂CH₂CH₂CH₂—,—CH₂CH₂CH₂CH₂CH₂CH₂CH₂CH₂— or substituted equivalents thereof. Thealkylene group may be unsubstituted or substituted by one or moresubstituents.

The term “cycloalkyl” refers to a saturated hydrocarbon ring system. Theterm “C₃-C₈ cycloalkyl” refers to a saturated hydrocarbon ring systemcontaining 3, 4, 5, 6, 7 or 8 carbon atoms. The ring system may be asingle ring or a bi-cyclic or tri-cyclic ring system. Where the ringsystem is bicyclic one of the rings may be an aromatic ring, for exampleas in indane. The term “cycloalkyl” may refer to, for example,cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl,cyclooctyl, and indane. The cycloalkyl group may be substituted with oneor more substituents.

The term “haloalkyl” refers to a linear or branched hydrocarbon chainwhich is substituted with at least one halogen atom which areindependently selected at each occurrence from fluorine, chlorine,bromine and iodine. For example, the term “C₁-C₆ haloalkyl” refers to alinear or branched hydrocarbon chain containing 1, 2, 3, 4, 5 or 6carbon atoms. The halogen atom may be at substituted at any position onthe hydrocarbon chain. The term “C₁-C₆ haloalkyl” may refer to, forexample, fluoromethyl, trifluoromethyl, chloromethyl, fluoroethyl,trifluoroethyl, chloroethyl, trichloroethyl (such as1,2,2-trichloroethyl and 2,2,2-trichloroethyl), fluoropropyl andchloropropyl. The haloalkyl group may be substituted with one or moresubstituents.

The term “alkenyl” refers to a linear or branched hydrocarbon chaincontaining at least one carbon-carbon double bond and having at leasttwo carbon atoms. The term “C₂-C₆ alkenyl” refers to a linear orbranched hydrocarbon chain containing at least one carbon-carbon doublebond and having 2, 3, 4, 5 or 6 carbon atoms. The double bond or doublebonds may be E or Z isomers. The double bond may be present at anypossible position of the hydrocarbon chain. The term “C₂-C₆ alkenyl” mayrefer to, for example, ethenyl, propenyl, butenyl, butadienyl, pentenyl,pentadienyl, hexenyl and hexadienyl. The alkenyl group may besubstituted or unsubstituted by one or more substituents.

The term “cycloalkenyl” refers to an unsaturated hydrocarbon ringsystem. The term “C₃-C₈ cycloalkenyl” refers to an unsaturatedhydrocarbon ring system containing 3, 4, 5, 6, 7 or 8 carbon atoms. Thering may contain more than one double bond. The term cycloalkenyl mayrefer to, for example cyclopropenyl, cyclobutenyl, cyclopentenyl,cyclopentadienyl, cyclohexenyl, cyclohexadienyl, cycloheptenyl,cycloheptadiene, cyclooctenyl and cycloocatadienyl. The cycloalkenylgroup may be substituted with one or more substituents.

The term “alkynyl” refers to a linear or branched hydrocarbon chaincontain at least one carbon-carbon triple bond and having at least twocarbon atoms. The term “C₂-C₆ alkynyl” refers to a linear or branchedhydrocarbon chain containing at least one carbon-carbon triple bond andhaving 2, 3, 4, 5 or 6 carbon atoms. The triple bond or triple bonds maybe present at any possible position of the hydrocarbon chain. The term“C₂-C₆ alkynyl” may refer to, for example, ethynyl, propynyl, butynyl,pentynyl and hexynyl. The alkynyl group may be unsubstituted orsubstituted by one or more substituents.

The term “heteroalkyl” refers to a linear or branched hydrocarbon chaincontaining at least one heteroatom selected from N, O and S which ispositioned between any possible carbon atom in the chain or at the endof the chain. The term “C₁-C₆ heteroalkyl” refers to a linear orbranched hydrocarbon chain containing 1, 2, 3, 4, 5, or 6 carbon atomsand at least one heteroatom selected from N, O and S which is positionedbetween any possible carbon atom in the chain or at the end of thechain. The heteroalkyl may be attached to another group by theheteroatom or the carbon atom. The term “C₁-C₆ heteroalkyl” may referto, for example, —CH₂NHCH₃, —NHCH₂CH₃ and —CH₂CH₂NH₂. The heteroalkylgroup may be unsubstituted or substituted by one or more substituents.

The term “heterocycloalkyl” refers to a saturated hydrocarbon ringsystem containing at least one heteroatom within the ring systemselected from N, O and S. The term “5- to 8-membered heterocycloalkyl”refers to a saturated hydrocarbon ring with 5, 6, 7, 8, 9 or 10 atomsselected from carbon, N, O and S, at least one being a heteroatom. The“heterocycloalkyl” group may also be denoted as a “3 to 10 memberedheterocycloalkyl” which is also a ring system containing 3, 4, 5, 6, 7,8, 9 or 10 atoms, at least one being a heteroatom. The ring system maybe a single ring or a bi-cyclic or tri-cyclic ring system. Bicyclicsystems may be spiro-fused, i.e. where the rings are linked to eachother through a single carbon atom; vicinally fused, i.e. where therings are linked to each other through two adjacent carbon or nitrogenatoms; or they may be share a bridgehead, i.e. the rings are linked toeach other two non-adjacent carbon or nitrogen atoms. Where the ringsystem is bicyclic one of the rings may be an aromatic ring, for exampleas in chromane. The “heterocycloalkyl” may be bonded to the rest of themolecule through any carbon atom or heteroatom. The “heterocycloalkyl”may have one or more, e.g. one or two, bonds to the rest of themolecule: these bonds may be through any of the atoms in the ring. Forexample, the “heterocycloalkyl” may be oxirane, aziridine, azetidine,oxetane, tetrahydrofuran, pyrrolidine, imidazolidine, succinimide,pyrazolidine, oxazolidine, isoxazolidine, thiazolidine, isothiazolidine,piperidine, morpholine, thiomorpholine, piperazine, tetrahydropyran, andchromane.

The term “heterocycloalkenyl” refers to an unsaturated hydrocarbon ringsystem containing at least one heteroatom selected from N, O or S. Theterm “C₃-C₈ heterocycloalkenyl” refers to an unsaturated hydrocarbonring system containing 3, 4, 5, 6, 7 or 8 carbon atoms and at least oneheteroatom selected from N, O or S. There may be more than one doublebond present. The double bond will typically be between two carbon atomsbut may be between a carbon atom and a nitrogen atom. There may also bemore than 1 heteroatom present. For example, there may be 1, 2 or 3heteroatoms present. The ring system may be a single ring or a bi-cyclicor tri-cyclic ring system. Where the ring system is bicyclic one of therings may be an aromatic ring, for example as in indoline anddihydrobenzofuran. The heterocycloalkenyl may be attached to anothergroup by any carbon or heteroatom. The term heterocycloalkenyl may referto, for example tetrahydropyridine, dihydropyran, dihydrofuran,pyrroline, dihydrobenzofuran, dihydrobenzothiophene and indoline. Theheterocycloalkenyl group may be substituted with one or moresubstituents.

The term “aryl” refers to an aromatic hydrocarbon ring system whichsatisfies Huckel's rule for aromaticity or that contains a ring systemwhich satisfies Huckel's rule for aromaticity. As such an aryl group maybe a single ring or a bi-cyclic or tri-cyclic ring system. The term“aryl” may refer to, for example, phenyl, naphthyl, indane, tetralin andanthracene. The aryl group may be unsubstituted or substituted with oneor more substituents. Any aryl group may be a phenyl ring.

The term “heteroaryl” refers to an aromatic hydrocarbon ring system withat least one heteroatom selected from N, O or S which satisfies Huckel'srule for aromaticity or a ring system that contains a heteroatom and anaromatic hydrocarbon ring. The heteroaryl may be a single ring system ora fused ring system. The term “5-, 6-, 9- or 10-membered heteroaryl”refers to an aromatic ring system within 5, 6, 9, or 10 members selectedfrom carbon, N, O or S either in a single ring or a bicyclic ringsystem. The term heteroaryl may refer to, for example, imidazole,thiazole, oxazole, isothiazole, isoxazole, triazole, tetraazole,thiophene, furan, thianthrene, pyrrole, benzimidazole, pyrazole,pyrazine, pyridine, pyrimidine, indole, isoindole, quinolone, andisoquinoline.

The term “alkoxy” refers to an alkyl group which is linked to anothergroup by oxygen. The alkyl group may be linear or branched. The term“C₁-C₆ alkoxy” refers to an alkyl group containing 1, 2, 3, 4, 5 or 6carbon atoms which is linked to another group by oxygen. The alkyl groupmay be, for example, methyl, ethyl, n-propyl, iso-propyl, n-butyl,sec-butyl, tert-butyl, n-pentyl and n-hexyl. The term “C₁-C₆ alkoxy” mayrefer to, for example, methoxy, ethoxy, n-propoxy, iso-propoxy,n-butoxy, sec-butoxy, tert-butoxy, n-pentoxy and n-hexoxy. The alkylgroup may be substituted or unsubstituted by one or more substituents.

A bond terminating in a “

” means that the bond is connected to another group that is not shown. Abond terminating inside a cyclic structure and not terminating at anatom of the ring structure represents that the bond may be connected toany of the atoms in the ring structure where allowed by valency.

Where a group is substituted, it may be substituted at any point on thegroup where chemically possible and consistent with valencyrequirements. The group may be substituted by one or more substituents.For example, the group may be substituted with 1, 2, 3 or 4substituents. Where there are two or more substituents, the substituentsmay be the same or different. Substituent(s) may be, for example, halo,CN, nitro, oxo, C₁-C₆-alkyl, C₂-C₆-alkynyl, C₂-C₆-alkenyl,C₁-C₆-haloalkyl, OR^(a), NR^(a)R^(b), SR^(a), C(O)OR^(a),C(O)NR^(a)R^(a), halo, cyano, nitro, C(O)R^(a), S(O)₂OR^(a), S(O)₂R^(a)and S(O)₂NR^(a)R^(a); wherein R^(a) is independently at each occurrenceselected from: H and C₁-C₆-alkyl; and R^(b) is independently at eachoccurrence selected from: H, C₁-C₆-alkyl, C(O)C₁-C₆-alkyl andS(O)₂—C₁-C₆-alkyl.

If chemically possible to do so, a cyclic substituent may be substitutedon a group so as to form a spiro-cycle.

Substituents are only present at positions where they are chemicallypossible, the person skilled in the art being able to decide (eitherexperimentally or theoretically) without inappropriate effort whichsubstitutions are chemically possible and which are not.

Ortho, meta and para substitution are well understood terms in the art.For the absence of doubt, “ortho” substitution is a substitution patternwhere adjacent carbons possess a substituent, whether a simple group,for example the fluoro group in the example below, or other portions ofthe molecule, as indicated by the bond ending in “

”.

“Meta” substitution is a substitution pattern where two substituents areon carbons one carbon removed from each other, i.e with a single carbonatom between the substituted carbons. In other words there is asubstituent on the second atom away from the atom with anothersubstituent. For example the groups below are meta substituted.

“Para” substitution is a substitution pattern where two substituents areon carbons two carbons removed from each other, i.e with two carbonatoms between the substituted carbons. In other words there is asubstituent on the third atom away from the atom with anothersubstituent. For example the groups below are para substituted.

The cation of formula (I) will be associated with a pharmaceuticallyacceptable anionic counter ion for administration to a subject.Nevertheless, where either the cation of formula (I) or the anioniccounter ion comprise either basic or acidic groups, those groups maythemselves be protonated or deprotonated and associated with anappropriate counter ion.

Suitable acid addition salts are formed from acids which form non-toxicsalts, for example, acetate, aspartate, benzoate, besylate,bicarbonate/carbonate, bisulphate/sulphate, borate, camsylate, citrate,edisylate, esylate, formate, fumarate, gluceptate, gluconate,glucuronate, hexafluorophosphate, hibenzate, hydrochloride/chloride,hydrobromide/bromide, hydroiodide/iodide, isethionate, lactate, malate,maleate, malonate, mesylate, methylsulphate, naphthylate,1,5-naphthalenedisulfonate, 2-napsylate, nicotinate, nitrate, orotate,oxalate, palmitate, pamoate, phosphate/hydrogen phosphate/dihydrogenphosphate, saccharate, stearate, succinate, tartrate, tosylate andtrifluoroacetate salts.

Suitable base salts are formed from bases which form non-toxic salts,for example include the aluminium, arginine, benzathine, calcium,choline, diethylamine, diolamine, glycine, lysine, magnesium, meglumine,olamine, potassium, sodium, tromethamine and zinc salts. Hemisalts ofacids and bases may also be formed, for example, hemisulphate andhemicalcium salts. A review of suitable salts can be found in “Handbookof Pharmaceutical Salts: Properties, Selection, and Use” by Stahl andWermuth (Wiley-VCH, Weinheim, Germany, 2002).

The salt may be an acid addition salt.

The salts may be formate or hydrochloride.

Pharmaceutically acceptable salts of compounds of formula (I) may beprepared by one or more of the following methods:

(i) reacting the compound of formula (I) with the desired acid or base;(ii) removing an acid- or base-labile protecting group from a suitableprecursor of the compound of formula (I) or by ring-opening a suitablecyclic precursor, for example, a lactone or lactam, using the desiredacid or base; or(iii) converting one salt of the compound of formula (I) to another byreaction with an appropriate acid or base or by means of a suitable ionexchange column.

The reactions above are typically carried out in solution and theresulting salt may precipitate out and be collected by filtration or maybe recovered by evaporation of the solvent. The degree of ionisation inthe resulting salt may vary from completely ionised to almostnon-ionised.

The compounds may exist in both unsolvated and solvated forms. The term‘solvate’ is used herein to describe a molecular complex comprising thecompound of the invention and a stoichiometric amount of one or morepharmaceutically acceptable solvent molecules, for example, ethanol. Theterm ‘hydrate’ is employed when said solvent is water.

Complexes are contemplated, such as clathrates, drug-host inclusioncomplexes wherein, in contrast to the aforementioned solvates, the drugand host are present in stoichiometric or non-stoichiometric amounts.Complexes of the drug containing two or more organic and/or inorganiccomponents which may be in stoichiometric or non-stoichiometric amountsare also contemplated. The resulting complexes may be ionised, partiallyionised, or non-ionised. A review of such complexes is found in J PharmSci, 64 (8), 1269-1288 by Haleblian (August 1975).

Compounds and salts described in this specification may beisotopically-labelled (or “radio-labelled”). Accordingly, one or moreatoms are replaced by an atom having an atomic mass or mass numberdifferent from the atomic mass or mass number typically found in nature.Examples of radionuclides that may be incorporated include ²H (alsowritten as “D” for deuterium), ³H (also written as “T” for tritium),¹¹C, ¹³C, ¹⁴C, ¹⁵O, ¹⁷O, ¹⁸O, ¹⁸F and the like. The radionuclide that isused will depend on the specific application of that radio-labelledderivative. For example, for in vitro competition assays, ³H or ¹⁴C areoften useful. For radio-imaging applications, ¹¹C or ¹⁸F are oftenuseful. In some embodiments, the radionuclide is ³H. In someembodiments, the radionuclide is ¹⁴C. In some embodiments, theradionuclide is ¹¹C. And in some embodiments, the radionuclide is ¹⁸F.

Hereinafter all references to compounds of any formula includereferences to salts, solvates and complexes thereof and to solvates andcomplexes of salts thereof.

The compounds include a number of formula as herein defined, includingall polymorphs and crystal habits thereof, prodrugs and isomers thereof(including optical, geometric and tautomeric isomers) as hereinafterdefined and isotopically-labelled compounds of the invention.

Before purification, the compounds may exist as a mixture of enantiomersdepending on the synthetic procedure used. The enantiomers can beseparated by conventional techniques known in the art. Thus thecompounds cover individual enantiomers as well as mixtures thereof.

For some of the steps of the process of preparation of the compounds offormula (I), it may be necessary to protect potential reactive functionsthat are not wished to react, and to cleave said protecting groups inconsequence. In such a case, any compatible protecting radical can beused. In particular methods of protection and deprotection such as thosedescribed by T. W. Greene (Protective Groups in Organic Synthesis, A.Wiley-Interscience Publication, 1981) or by P. J. Kocienski (Protectinggroups, Georg Thieme Verlag, 1994), can be used. All of the abovereactions and the preparations of novel starting materials used in thepreceding methods are conventional and appropriate reagents and reactionconditions for their performance or preparation as well as proceduresfor isolating the desired products will be well-known to those skilledin the art with reference to literature precedents and the examples andpreparations hereto.

Also, the compounds as well as intermediates for the preparation thereofcan be purified according to various well-known methods, such as forexample crystallization or chromatography.

The method of treatment or the compound for use in the treatment ofsolid tumours, leukaemia, lymphoma, sarcoma, or carcinoma as definedhereinbefore may be applied as a sole therapy or be a combinationtherapy with an additional active agent.

The method of treatment or the compound for use in the treatment ofsolid tumours, leuekaemia, lymphoma, sarcoma, or carcinoma may involve,in addition to the compound of the invention, conventional surgery orradiotherapy or chemotherapy. Such chemotherapy may include one or moreof the following categories of anti-cancer agents:

(i) antiproliferative/antineoplastic drugs and combinations thereof,such as alkylating agents (for example cis-platin, oxaliplatin,carboplatin, cyclophosphamide, nitrogen mustard, bendamustin, melphalan,chlorambucil, busulphan, temozolamide and nitrosoureas); antimetabolites(for example gemcitabine and antifolates such as fluoropyrimidines like5-fluorouracil and tegafur, raltitrexed, methotrexate, pemetrexed,cytosine arabinoside, and hydroxyurea); antibiotics (for exampleanthracyclines like adriamycin, bleomycin, doxorubicin, daunomycin,epirubicin, idarubicin, mitomycin-C, dactinomycin and mithramycin);antimitotic agents (for example vinca alkaloids like vincristine,vinblastine, vindesine and vinorelbine and taxoids like taxol andtaxotere and polokinase inhibitors); proteasome inhibitors, for examplecarfilzomib and bortezomib; interferon therapy; and topoisomeraseinhibitors (for example epipodophyllotoxins like etoposide andteniposide, amsacrine, topotecan, mitoxantrone and camptothecin);(ii) cytostatic agents such as antiestrogens (for example tamoxifen,fulvestrant, toremifene, raloxifene, droloxifene and iodoxyfene),antiandrogens (for example bicalutamide, flutamide, nilutamide andcyproterone acetate), LHRH antagonists or LHRH agonists (for examplegoserelin, leuprorelin and buserelin), progestogens (for examplemegestrol acetate), aromatase inhibitors (for example as anastrozole,letrozole, vorazole and exemestane) and inhibitors of 5α-reductase suchas finasteride;(iii) anti-invasion agents, for example dasatinib and bosutinib(SKI-606), and metalloproteinase inhibitors, inhibitors of urokinaseplasminogen activator receptor function or antibodies to Heparanase;(iv) inhibitors of growth factor function: for example such inhibitorsinclude growth factor antibodies and growth factor receptor antibodies,for example the anti-erbB2 antibody trastuzumab [Herceptin™], theanti-EGFR antibody panitumumab, the anti-erbB1 antibody cetuximab,tyrosine kinase inhibitors, for example inhibitors of the epidermalgrowth factor family (for example EGFR family tyrosine kinase inhibitorssuch as gefitinib, erlotinib and6-acrylamido-/V-(3-chloro-4-fluorophenyl)-7-(3-morpholinopropoxy)-quinazolin-4-amine(Cl 1033), erbB2 tyrosine kinase inhibitors such as lapatinib);inhibitors of the hepatocyte growth factor family; inhibitors of theinsulin growth factor family; modulators of protein regulators of cellapoptosis (for example Bcl-2 inhibitors); inhibitors of theplatelet-derived growth factor family such as imatinib and/or nilotinib(AMN107); inhibitors of serine/threonine kinases (for example Ras/Rafsignalling inhibitors such as farnesyl transferase inhibitors, forexample sorafenib, tipifarnib and lonafarnib), inhibitors of cellsignalling through MEK and/or AKT kinases, c-kit inhibitors, abl kinaseinhibitors, PI3 kinase inhibitors, Plt3 kinase inhibitors, CSF-1R kinaseinhibitors, IGF receptor, kinase inhibitors; aurora kinase inhibitorsand cyclin dependent kinase inhibitors such as CDK2 and/or CDK4inhibitors;(v) antiangiogenic agents such as those which inhibit the effects ofvascular endothelial growth factor, for example the anti-vascularendothelial cell growth factor antibody bevacizumab (Avastin™);thalidomide; lenalidomide; and for example, a VEGF receptor tyrosinekinase inhibitor such as vandetanib, vatalanib, sunitinib, axitinib andpazopanib;(vi) gene therapy approaches, including for example approaches toreplace aberrant genes such as aberrant p53 or aberrant BRCA1 or BRCA2;(vii) immunotherapy approaches, including checkpoint inhibitors oftargets such as PD-1, PD-L1 and CTCLA-4 for example antibody therapysuch as alemtuzumab, rituximab, ibritumomab tiuxetan (Zevalin®),pembrolizumab and ofatumumab; interferons such as interferon α;interleukins such as IL-2 (aldesleukin); interleukin inhibitors forexample IRAK4 inhibitors; cancer vaccines including prophylactic andtreatment vaccines such as HPV vaccines, for example Gardasil, Cervarix,Oncophage and Sipuleucel-T (Provenge); and toll-like receptor modulatorsfor example TLR-7 or TLR-9 agonists; and(viii) cytotoxic agents for example fludaribine (fludara), cladribine,pentostatin (Nipent™);(ix) steroids such as corticosteroids, including glucocorticoids andmineralocorticoids, for example aclometasone, aclometasone dipropionate,aldosterone, amcinonide, beclomethasone, beclomethasone dipropionate,betamethasone, betamethasone dipropionate, betamethasone sodiumphosphate, betamethasone valerate, budesonide, clobetasone, clobetasonebutyrate, clobetasol propionate, cloprednol, cortisone, cortisoneacetate, cortivazol, deoxycortone, desonide, desoximetasone,dexamethasone, dexamethasone sodium phosphate, dexamethasoneisonicotinate, difluorocortolone, fluclorolone, flumethasone,flunisolide, fluocinolone, fluocinolone acetonide, fluocinonide,fluocortin butyl, fluorocortisone, fluorocortolone, fluocortolonecaproate, fluocortolone pivalate, fluorometholone, fluprednidene,fluprednidene acetate, flurandrenolone, fluticasone, fluticasonepropionate, halcinonide, hydrocortisone, hydrocortisone acetate,hydrocortisone butyrate, hydrocortisone aceponate, hydrocortisonebuteprate, hydrocortisone valerate, icomethasone, icomethasone enbutate,meprednisone, methylprednisolone, mometasone paramethasone, mometasonefuroate monohydrate, prednicarbate, prednisolone, prednisone,tixocortol, tixocortol pivalate, triamcinolone, triamcinolone acetonide,triamcinolone alcohol and their respective pharmaceutically acceptablederivatives. A combination of steroids may be used, for example acombination of two or more steroids mentioned in this paragraph;(x) targeted therapies, for example PI3K5 inhibitors, for exampleidelalisib and perifosine.

Such combination treatment may be achieved by way of the simultaneous,sequential or separate dosing of the individual components of thetreatment. Such combination products may be administered so that thecombination is provided in a therapeutically effective amount, forexample the compounds of this invention may be administered within atherapeutically effective dosage range described herein and the otherpharmaceutically-active agent may be administered in an amount of lessthan or within its approved dosage range.

According to a further aspect of the invention there is provided apharmaceutical product comprising a compound of the first aspect of theinvention, or a pharmaceutically acceptable salt thereof as definedherein and an additional active agent. The additional active agent maybe a cancer therapy as defined hereinbefore for the combinationtreatment of cancer.

According to a further aspect of the invention there is provided amethod of treating cancer comprising administering a therapeuticallyeffective amount of a compound of the invention, or a pharmaceuticallyacceptable salt thereof simultaneously, sequentially or separately withan additional anti-cancer agent, as defined hereinbefore, to a patientin need thereof.

According to a further aspect of the invention there is provided acompound of the invention, or a pharmaceutically acceptable salt thereoffor use simultaneously, sequentially or separately with an additionalanti-cancer agent as defined herein, in the treatment of cancer.

According to another aspect of the invention there is provided a use ofthe compound of the invention in combination with an anti-cancer agent,such as those hereinbefore described. The compound of formula (I) may beused simultaneously, sequentially or separately with the additionalanti-cancer agent. The use may be in a single combination productcomprising the compound of the invention and the anti-cancer agent. Theadditional anti-cancer agent may be a further compound of the firstaspect of the invention.

According to a further aspect there is provided a method of providing acombination product, wherein the method comprises providing a compoundof the invention simultaneously, sequentially or separately with ananti-cancer agent, as defined hereinbefore. The method may comprisecombining the compound of the invention and the anti-cancer agent in asingle dosage form. Alternatively the method may comprise providing theanti-cancer agent as separate dosage forms.

Compounds of the invention may exist in a single crystal form or in amixture of crystal forms or they may be amorphous. Thus, compounds ofthe invention intended for pharmaceutical use may be administered ascrystalline or amorphous products. They may be obtained, for example, assolid plugs, powders, or films by methods such as precipitation,crystallization, freeze drying, or spray drying, or evaporative drying.Microwave or radio frequency drying may be used for this purpose.

For the above-mentioned compounds of the invention the dosageadministered will, of course, vary with the compound employed, the modeof administration, the treatment desired and the disorder indicated. Forexample, if the compound of the invention is administered orally, thenthe daily dosage of the compound of the invention may be in the rangefrom 0.01 micrograms per kilogram body weight (μg/kg) to 100 milligramsper kilogram body weight (mg/kg).

A compound of the invention, or pharmaceutically acceptable saltthereof, may be used on their own but will generally be administered inthe form of a pharmaceutical composition in which the compounds of theinvention, or pharmaceutically acceptable salt thereof, is inassociation with a pharmaceutically acceptable adjuvant, diluent orcarrier. Conventional procedures for the selection and preparation ofsuitable pharmaceutical formulations are described in, for example,“Pharmaceuticals—The Science of Dosage Form Designs”, M. E. Aulton,Churchill Livingstone, 1988.

Depending on the mode of administration of the compounds of theinvention, the pharmaceutical composition which is used to administerthe compounds of the invention will preferably comprise from 0.05 to 99%w (percent by weight) compounds of the invention, more preferably from0.05 to 80% w compounds of the invention, still more preferably from0.10 to 70% w compounds of the invention, and even more preferably from0.10 to 50% w compounds of the invention, all percentages by weightbeing based on total composition.

The pharmaceutical compositions may be administered topically (e.g. tothe skin) in the form, e.g., of creams, gels, lotions, solutions,suspensions, or systemically, e.g. by oral administration in the form oftablets, capsules, syrups, powders or granules; or by parenteraladministration in the form of a sterile solution, suspension or emulsionfor injection (including intravenous, subcutaneous, intramuscular,intravascular or infusion); by rectal administration in the form ofsuppositories; or by inhalation in the form of an aerosol.

For oral administration the compounds of the invention may be admixedwith an adjuvant or a carrier, for example, lactose, saccharose,sorbitol, mannitol; a starch, for example, potato starch, corn starch oramylopectin; a cellulose derivative; a binder, for example, gelatine orpolyvinylpyrrolidone; and/or a lubricant, for example, magnesiumstearate, calcium stearate, polyethylene glycol, a wax, paraffin, andthe like, and then compressed into tablets. If coated tablets arerequired, the cores, prepared as described above, may be coated with aconcentrated sugar solution which may contain, for example, gum arabic,gelatine, talcum and titanium dioxide. Alternatively, the tablet may becoated with a suitable polymer dissolved in a readily volatile organicsolvent.

For the preparation of soft gelatine capsules, the compounds of theinvention may be admixed with, for example, a vegetable oil orpolyethylene glycol. Hard gelatine capsules may contain granules of thecompound using either the above-mentioned excipients for tablets. Alsoliquid or semisolid formulations of the compound of the invention may befilled into hard gelatine capsules. Liquid preparations for oralapplication may be in the form of syrups or suspensions, for example,solutions containing the compound of the invention, the balance beingsugar and a mixture of ethanol, water, glycerol and propylene glycol.Optionally such liquid preparations may contain colouring agents,flavouring agents, sweetening agents (such as saccharine), preservativeagents and/or carboxymethylcellulose as a thickening agent or otherexcipients known to those skilled in art.

For intravenous (parenteral) administration the compounds of theinvention may be administered as a sterile aqueous or oily solution.

The size of the dose for therapeutic purposes of compounds of theinvention will naturally vary according to the nature and severity ofthe conditions, the age and sex of the animal or patient and the routeof administration, according to well-known principles of medicine.

Dosage levels, dose frequency, and treatment durations of compounds ofthe invention are expected to differ depending on the formulation andclinical indication, age, and co-morbid medical conditions of thepatient.

Throughout the description and claims of this specification, the words“comprise” and “contain” and variations of them mean “including but notlimited to”, and they are not intended to (and do not) exclude othermoieties, additives, components, integers or steps. Throughout thedescription and claims of this specification, the singular encompassesthe plural unless the context otherwise requires. In particular, wherethe indefinite article is used, the specification is to be understood ascontemplating plurality as well as singularity, unless the contextrequires otherwise.

Features, integers, characteristics, compounds, chemical moieties orgroups described in conjunction with a particular aspect, embodiment orexample of the invention are to be understood to be applicable to anyother aspect, embodiment or example described herein unless incompatibletherewith. All of the features disclosed in this specification(including any accompanying claims, abstract and drawings), and/or allof the steps of any method or process so disclosed, may be combined inany combination, except combinations where at least some of suchfeatures and/or steps are mutually exclusive. The invention is notrestricted to the details of any foregoing embodiments. The inventionextends to any novel one, or any novel combination, of the featuresdisclosed in this specification (including any accompanying claims,abstract and drawings), or to any novel one, or any novel combination,of the steps of any method or process so disclosed.

The reader's attention is directed to all papers and documents which arefiled concurrently with or previous to this specification in connectionwith this application and which are open to public inspection with thisspecification, and the contents of all such papers and documents areincorporated herein by reference.

Methods for Synthesizing Compounds

Certain ions of the invention can be synthesised according to oranalogously to methods described in the General Schemes below and/byother techniques known to those of ordinary skill in the art. Certainions of the invention can be synthesised according to or analogously tothe methods described in the Examples.

Certain compounds of formula (II) can be made by Scheme A.

Amide bond formation between N-desmethyl-azithromycin (1) andphosphonium carboxylic acid (2) can deliver phosphonium amide (3). Thereaction can be effected by standard peptide coupling reagents, such asHATU or TATU or 1,3-dicyclohexylcarboiimide (DCC) in the presence ofHOAT, in the presence of a base, such as N,N-diisopropylethylamine(DIEA), in an organic solvent, such as DCM, at a temperature of 25 to40° C. Phosphonium carboxylic acid (2) can be prepared from reaction ofhalide (4) (X═Cl or Br) with phosphine PR^(1a)R^(1b)R^(1c) (7). Thereaction can be accomplished by heating in an organic solvent, such asMeCN at a temperature from 50 to 80° C.

Certain compounds of formula (II) can be made by Scheme B.

Reaction of N-desmethyl-azithromycin (1) with halo sulphonic acid (5)(where X═C₁ or Br) can furnish halo sulphonamide (6). The reaction canbe performed using SOCl₂ in an organic solvent, such as DMF or DCM, at atemperature from 20 to 60° C. Reaction of phosphine (7) with halosulphonamide (6) can deliver phosphonium sulphonamide (8). The reactioncan be accomplished by heating in an organic solvent, such as MeCN at atemperature from 50 to 80° C.

Certain compounds of formula (II) can be made by Scheme C.

Reaction of N-desmethyl-azithromycin (1) with phosphoniumchlorocarbonate (9) can furnish phosphonium carbamate (10). The reactioncan be accomplished in the presence of a base, such as pyridine, in anorganic solvent, such as DCM, at a temperature from 0° C. to roomtemperature. Phosphonium chloroacetate (9) can be synthesised fromphosphonium alcohol (11) through treatment with triphosgene in thepresence of a base, such as N,N-diisopropylethylamine (DIEA), in anorganic solvent, such as THF, at a temperature from −5 to 5° C.Phosphonium alcohol (11) can be synthesised from halo alcohol (12)(where X═C₁ or Br) and phosphine (7). The reaction can be accomplishedby heating in an organic solvent, such as MeCN, at a temperature from 50to 80° C.

Certain compounds of formula (II) can be made by Scheme D.

Reductive amination of N-desmethyl-azithromycin (1) with aldehyde (13),can furnish amine (14), where LG represents a leaving group, such as Cl,Br, I, tosyl or mesylate and L^(2a) is one carbon shorter than L². Thereaction can be accomplished using an appropriate reducing agent, suchas NaBH₃CN in the presence of a source of acid, such as CH₃COOH, in anorganic solvent, such as DMF, at a temperature of 25 to 80° C.Displacement of the LG in (14) with phosphine (7) can deliverphosphonium amine (15). The reaction can be performed in an organicsolvent such as MeCN, at a temperature of 50 to 90° C. As an adaptationaddition of a metal iodide, such as NaI or KI in the cases where the LGis not I can facilitate formation of the I in situ as a potentially moreeffective LG.

Certain compounds of formula (II) can be made by Scheme E.

Reaction of phosphonium amides (3) with Ac₂O can selectively acylate thehydroxy group in the desoaminyl sugar to give the acylated phosphoniumamide (16). The reaction can be accomplished in the presence of a base,such as Et₃N, in the presence of an organic solvent, such as DCM, atroom temperature. Sequential treatment of (16) with carbonyldimidazole(CDI) in the presence of a base, such as Et₃N, at a temperature of 25 to70° C., followed by treatment with a strong base, such as NaOH, in anorganic solvent, such as THF, at a temperature of 0 to 25° C., followedby heating in MeOH, at a temperature of 40 to 65° C. can furnishphosphonium carbonate (17).

Certain compounds of formula (VIIb) can be made by Scheme F.

Reaction of (18) (prepared by the procedure described in EuropeanJournal of Medicinal Chemistry 40, 2011, 5196) with a strong base, suchas NaOH, in the presence of a solvent, such as THF, at a temperature of0 to 25° C., followed by ring opening of the carbonate with phosphoniumamine (20) can deliver carbamate (19). The carbonate ring openingreaction can be accomplished in the presence of a non-nucleophilic base,such as DBU, in pyridine at room temperature. Removal of the acetate inthe desoaminyl sugar can be accomplished by heating in MeOH, at atemperature of 40 to 65° C. to furnish phosphonium carbamate (21).

Phosphonium amine (20) can be furnished by Scheme G.

Reaction of halo isoindoline-1,3-dione (22) (where X═C₁ or Br) withphosphine (7) can deliver phosphonium isoindoline-1,3-dione (23). Thereaction can be performed by heating in an organic solvent, such asMeCN, at a temperature from 50 to 80° C. Reaction of the phosphoniumisoindoline-1,3-dione (23) with hydrazine hydrate in an alcoholicsolvent, such as EtOH, at a temperature from 50 to 75° C. can furnishphosphonium amine (20).

Certain compounds of formula (II) can be made by Scheme H.

Removal of the cladinosyl sugar in phosphonium amide (3) to give (24)can be accomplished by the treatment of a mineral acid, such as HCl, ina solvent such as MeOH, at room temperature.

Replacement of amine (1) in Scheme A with amine (25) (prepared asdescribed in WO2011116312) can give phosphonium amide (26).

EXPERIMENTAL Analytical Methods

NMR spectra were obtained on a 400 MHz Bruker AV III

UPLC/MS was carried out using a Waters Acquity QDa mass detector andMethods A, G or Waters SQ mass detector and Methods B, C, D, E, F, H

Method A

Column: Waters Acquity UPLC CSH C18, 1.7 μm, 2.1×30 mm; Gradient Eluent:5-95% MeCN/H₂O containing 0.1% HCOOH; Time: 0-10 min

Method B

Column: Waters Acquity UPLC CSH C18, 1.7 μm, 2.1×50 mm; Gradient Eluent:2-98% MeCN/H₂O containing 0.02% HCOOH; Time: 0-4.5 min

Method C

Column: Waters Acquity UPLC CSH C18, 1.7 μm, 2.1×50 mm; Gradient Eluent:2-98% MeCN containing 0.035% TFA/H₂O containing 0.05% TFA; Time: 0-4.5min

Method D

Column: Waters Acquity UPLC CSH C18, 1.7 μm, 2.1×100 mm; GradientEluent: 2-98% MeCN containing 0.035% TFA/H₂O containing 0.05% TFA; Time:0-15 min

Method E

Column: Waters Acquity UPLC BEH C18, 1.7 μm, 2.1×30 mm; Gradient Eluent:5-95% MeCN/H₂O containing 10 mM (NH₄)₂CO₃; Time: 0-3 min

Method F

Column: Waters Acquity UPLC CSH C18, 1.7 μm, 2.1×50 mm; Gradient Eluent:2-100% MeCN containing 0.035% TFA/H₂O containing 0.05% TFA; Time: 0-3min

Method G

Column: Waters Acquity UPLC BEH C18, 1.7 μm, 2.1×30 mm; Gradient Eluent:5-95% MeCN/H₂O containing 10 mM (NH₄)₂CO₃; Time: 0-3 min

Method H

Column: Waters Acquity UPLC BEH C18, 1.7 μm, 2.1×30 mm; Gradient Eluent:5-95% MeCN/H₂O containing 10 mM (NH₄)₂CO₃; Time: 0-15 min

Method I

Column: Waters Acquity UPLC BEH C18, 1.7 μm, 2.1×30 mm; Gradient Eluent:5-95% MeCN/H₂O containing 0.1% HCOOH; Time: 0-3 min

Preparative HPLC was carried out using a ZQ Mass Spectrometer and MethodA or Gilson PLC2020 and Method B,C

Method A

Waters X-Select Prep-C18, 5 μm, 19×50 mm eluting with MeCN/H₂O/0.1%HCOOH

Method B

Gemini NX-C18, 10 μm, 30×250 mm eluting with a mixture of MeCN/0.035%TFA and H₂O/0.05% TFA

Method C

Gemini NX-C18, 10 μm, 50×300 mm eluting with a mixture of MeCN/H₂O/0.1%TFA

General Protocol for Amide Coupling (Scheme A)

A1—To a stirred solution of N-desmethyl-azithromycin (1 equivalent), thecorresponding phosphonium acid (1.1 equivalents) and HATU (1.2equivalents) in DCM (10 mL) was added N,N-diisopropylethylamine (DIEA)(2 equivalents). After stirring at room temperature for 16 to 20 h thereaction mixture is concentrated, and the resulting residue diluted with7M NH₃ MeOH and stirred for a further 16 to 20 h. The reaction mixtureis then concentrated under reduced pressure and diluted with EtOAc. Theorganic is washed sequentially with saturated aqueous NaHCO₃, NH₄Cl andbrine. The organic is dried over Na₂SO₄ and concentrated under reducedpressure to give the crude product, which is purified by chromatographyto give the desired final product

A2—To a stirred solution of N-desmethyl-azithromycin (1 equivalent), thecorresponding phosphonium acid (1.1 equivalents) and HATU (1.2equivalents) in DCM (10 mL) was added N,N-diisopropylethylamine (DIEA)(2 equivalents). After stirring at room temperature for 16 to 20 h thereaction mixture is concentrated under reduced pressure and diluted withEtOAc. The organic is washed with saturated aqueous NaHCO₃, dried overNa₂SO₄ and concentrated under reduced pressure. The resulting residue isdiluted with MeOH and the stirring solution heated at 50° C. for 16 to20 h. On cooling the reaction is concentrated under reduced pressure anddiluted with EtOAc. The organic is washed sequentially with saturatedaqueous NaHCO₃, NH₄Cl and brine. The organic is dried over Na₂SO₄ andconcentrated under reduced pressure to give the crude product, which ispurified by chromatography. The organic is dried over Na₂SO₄ andconcentrated under reduced pressure to give the crude product, which ispurified by chromatography to give the desired final product.

General Protocol for Reductive Amination (Scheme D)

Step 1—To a stirred solution of N-desmethyl-azithromycin (1 equivalent),LG-L_(2a)-CHO (where LG presents a leaving group; L_(2a) is one carbonshorter than L2) (3.5 equivalents) and CH₃COOH (10 equivalents) in DMF(10 mL) is added NaBH₃CN (2 equivalents). The resulting mixture isheated at 75° C. for 90 min to 16 h. On cooling the reaction is quenchedwith H₂O, saturated aqueous NaHCO₃ and diluted with DCM. The aqueouslayer is extracted further with DCM and the combined organics washedwith saturated aqueous NaHCO₃ and brine. After drying over Na₂SO₄ theorganic is passed through a phase separator and concentrated underreduced pressure to give the crude product, which is purified bychromatography and used in step 2.

Step 2—The resulting purified product from step 1 is dissolved in MeCN(10 mL) and treated with the corresponding phosphine (3 equivalents) andNaI (3 equivalents). After thermal heating at 70 to 85° C. for 16 to 20h the reaction mixture is cooled to room temperature and concentratedunder reduced pressure. The resulting residue is purified bychromatography to give the final desired product. As an adaptation, thereaction can be conducted under microwave conditions at 100° C. for 2 to6 h.

General Protocol for Formation of Phosphonium Carboxylic Acids (2)

To a solution of HOOC-L²-(L³-L⁴)n-Br (1 equivalent) in MeCN (10 mL) isadded the phosphine (3 equivalents). After heating at 60° C. for 16 to20 h the reaction mixture is cooled to room temperature and concentratedunder reduced pressure. The resulting residue is purified bychromatography to give the desired phosphonium carboxylic acid product.

The following abbreviations have been used throughout the specificationin the examples and the description: dichloromethane (DCM),tetrahydrofuran (THF), dimethylformamide (DMF), dimethyl sulfoxide(DMSO), 4-dimethylaminopyridine (DMAP), N,N,diisopropylethylamine(DIEA), trifluoroacetic acid (TFA), 1,8-diazabicyclo[5.4.0]undec-7-ene(DBU), 1-hydroxy-7-azabenzotriazole (HOAT),0-(7-azabenzotriazole-1-yl)N,N,N′,N′-tetramethyluroniumtetrafluoroborate (TATU),(1-[Bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium3-oxid hexafluorophosphate) (HATU).

Example1—{14-[(2R,3S,4R,5R,8R,10R,11R,12S,13S,14R)-11-{[(2S,3R,4S,6R)-4-(dimethylamino)-3-hydroxy-6-methyloxan-2-yl]oxy}-2-ethyl-3,4,10-trihydroxy-13-{[(2R,4R,5S,6S)-5-hydroxy-4-methoxy-4,6-dimethyloxan-2-yl]oxy}-3,5,8,10,12,14-hexamethyl-15-oxo-1-oxa-6-azacyclopentadecan-6-yl]-14-oxotetradecyl}triphenylphosphoniumhexafluorophosphate(V)

Prepared following amide coupling procedure A1 using(13-carboxytridecyl)triphenylphosphonium bromide. Title compoundisolated as a white solid.

LC-MS (Method G) 1206 [M]⁺; RT 2.09 min

Example2—{10-[(2R,3S,4R,5R,8R,10R,11R,12S,13S,14R)-11-{[(2S,3R,4S,6R)-4-(dimethylamino)-3-hydroxy-6-methyloxan-2-yl]oxy}-2-ethyl-3,4,10-trihydroxy-13-{[(2R,4R,5S,6S)-5-hydroxy-4-methoxy-4,6-dimethyloxan-2-yl]oxy}-3,5,8,10,12,14-hexamethyl-15-oxo-1-oxa-6-azacyclopentadecan-6-yl]-10-oxodecyl}triphenylphosphoniumhexafluorophosphate(V)

Prepared following amide coupling procedure A1 using(9-carboxynonyl)triphenylphosphonium bromide. Title compound isolated asa white solid.

LC-MS (Method E) 1150.2 [M]⁺; RT 1.94 min

Example3—{9-[(2R,3S,4R,5R,8R,10R,11R,12S,13S,14R)-11-{[(2S,3R,4S,6R)-4-(dimethylamino)-3-hydroxy-6-methyloxan-2-yl]oxy}-2-ethyl-3,4,10-trihydroxy-13-{[(2R,4R,5S,6S)-5-hydroxy-4-methoxy-4,6-dimethyloxan-2-yl]oxy}-3,5,8,10,12,14-hexamethyl-15-oxo-1-oxa-6-azacyclopentadecan-6-yl]-9-oxononyl}triphenylphosphoniumhexafluorophosphate(V)

Prepared following amide coupling procedure A1 using(8-carboxyoctyl)triphenylphosphonium bromide. Title compound isolated asa white solid.

LC-MS (Method E) 978.2 [M-cladinosyl]⁺; RT 1.99 min

Example4—{7-[(2R,3S,4R,5R,8R,10R,11R,12S,13S,14R)-11-{[(2S,3R,4S,6R)-4-(dimethylamino)-3-hydroxy-6-methyloxan-2-yl]oxy}-2-ethyl-3,4,10-trihydroxy-13-{[(2R,4R,5S,6S)-5-hydroxy-4-methoxy-4,6-dimethyloxan-2-yl]oxy}-3,5,8,10,12,14-hexamethyl-15-oxo-1-oxa-6-azacyclopentadecan-6-yl]-7-oxoheptyl}triphenylphosphoniumhexafluorophosphate(V)

Prepared following amide coupling procedure A1 using(6-carboxyhexyl)triphenylphosphonium bromide. Title compound isolated asa white solid.

LC-MS (Method E) 1108 [M]⁺; RT 1.77 min

Example5—{12-[(2R,3S,4R,5R,8R,10R,11R,12S,13S,14R)-11-{[(2S,3R,4S,6R)-4-(dimethylamino)-3-hydroxy-6-methyloxan-2-yl]oxy}-2-ethyl-3,4,10-trihydroxy-13-{[(2R,4R,5S,6S)-5-hydroxy-4-methoxy-4,6-dimethyloxan-2-yl]oxy}-3,5,8,10,12,14-hexamethyl-15-oxo-1-oxa-6-azacyclopentadecan-6-yl]-12-oxododecyl}triphenylphosphoniumhexafluorophosphate(V)

Prepared following amide coupling procedure A1 using(11-carboxyundecyl)triphenylphosphonium bromide. Title compound isolatedas a white solid.

LC-MS (Method E) 1178 [M]⁺; RT 2.22 min

Example6—{6-[(2R,3S,4R,5R,8R,10R,11R,12S,13S,14R)-11-{[(2S,3R,4S,6R)-4-(dimethylamino)-3-hydroxy-6-methyloxan-2-yl]oxy}-2-ethyl-3,4,10-trihydroxy-13-{[(2R,4R,5S,6S)-5-hydroxy-4-methoxy-4,6-dimethyloxan-2-yl]oxy}-3,5,8,10,12,14-hexamethyl-15-oxo-1-oxa-6-azacyclopentadecan-6-yl]-6-oxohexyl}triphenylphosphoniumhexafluorophosphate(V)

Prepared following amide coupling procedure A1 using(5-carboxypentyl)triphenylphosphonium bromide. Title compound isolatedas a white solid.

LC-MS (Method G) 1094 [M]⁺; RT 1.59 min

Example7—{8-[(2R,3S,4R,5R,8R,10R,11R,12S,13S,14R)-11-{[(2S,3R,4S,6R)-4-(dimethylamino)-3-hydroxy-6-methyloxan-2-yl]oxy}-2-ethyl-3,4,10-trihydroxy-13-{[(2R,4R,5S,6S)-5-hydroxy-4-methoxy-4,6-dimethyloxan-2-yl]oxy}-3,5,8,10,12,14-hexamethyl-15-oxo-1-oxa-6-azacyclopentadecan-6-yl]octyl}triphenylphosphoniumiodide

Prepared following the general reductive amination procedure using8-chlorooctanal in step 1 and trlphenylphosphine in step 2 under thermalheating conditions. Title compound isolated as a white solid.

1H NMR (Method A) (DMSO-d₆): δ (delta) ppm 7.93-7.86 (m, 3H), 7.78 (m,12H), 4.87 (dd, J=10.2, 2.8 Hz, 1H), 4.78 (d, J=4.9 Hz, 1H), 4.42 (d,J=7.3 Hz, 1H), 4.32 (s, 1H), 4.22 (d, J=7.5 Hz, 1H), 4.13-4.03 (m, 2H),4.03-3.93 (m, 1H), 3.93-3.80 (d, 1H), 3.72-3.62 (m, 1H), 3.61-3.47 (m,4H), 3.22 (s, 3H), 3.08-2.99 (m, 1H), 2.91 (dd, J=9.4, 7.4 Hz, 1H),2.84-2.75 (m, 1H), 2.75-2.64 (m, 2H), 2.61-2.52 (m, 1H), 2.47-2.34 (m,3H), 2.29-2.16 (m, 7H), 2.12-1.99 (m, 1H), 1.97-1.91 (m, 1H), 1.91-1.81(m, 1H), 1.81-1.69 (m, 1H), 1.66-1.57 (m, 1H), 1.56-1.34 (m, 9H),1.30-1.23 (m, 3H), 1.21-1.04 (m, 20H), 1.02-0.92 (m, 9H), 0.85 (d, J=6.7Hz, 3H), 0.79 (t, J=7.4 Hz, 3H); 31P NMR (162 MHz, DMSO-d₆): δ (delta)ppm+24.07 (s); LC-MS (Method E) 1108 [M]⁺; RT 2.03 min

Example8—{6-[(2R,3S,4R,5R,8R,10R,11R,12S,13S,14R)-11-{[(2S,3R,4S,6R)-4-(dimethylamino)-3-hydroxy-6-methyloxan-2-yl]oxy}-2-ethyl-3,4,10-trihydroxy-13-{[(2R,4R,5S,6S)-5-hydroxy-4-methoxy-4,6-dimethyloxan-2-yl]oxy}-3,5,8,10,12,14-hexamethyl-15-oxo-1-oxa-6-azacyclopentadecan-6-yl]hexyl}triphenylphosphoniumiodide

Prepared following the general reductive amination procedure using6-chlorohexanal in step 1 and triphenylphosphine in step 2 under thermalheating conditions. Title compound isolated as a white solid.

LC-MS (Method G) 1080 [M]⁺; RT 2.00 min

Example9—{8-[(2R,3S,4R,5R,8R,10R,11R,12S,13S,14R)-11-{[(2S,3R,4S,6R)-4-(dimethylamino)-3-hydroxy-6-methyloxan-2-yl]oxy}-2-ethyl-3,4,10-trihydroxy-13-{[(2R,4R,5S,6S)-5-hydroxy-4-methoxy-4,6-dimethyloxan-2-yl]oxy}-3,5,8,10,12,14-hexamethyl-15-oxo-1-oxa-6-azacyclopentadecan-6-yl]-8-oxooctyl}triphenylphosphoniumhexafluorophosphate(V)

Prepared following amide coupling procedure A1 using(7-carboxyheptyl)triphenylphosphonium bromide. Title compound isolatedas a white solid.

LC-MS (Method E) 1122 [M]⁺; RT 1.59 min

Example10—{8-[(3aR,4R,7R,8S,9S,10R,11R,13R,16R,16aR)-10-{[(2S,3R,4S,6R)-4-(dimethylamino)-3-hydroxy-6-methyloxan-2-yl]oxy}-4-ethyl-11-hydroxy-8-{[(2R,4R,5S,6S)-5-hydroxy-4-methoxy-4,6-dimethyloxan-2-yl]oxy}-3a,7,9,11,13,16-hexamethyl-2,6-dioxo-tetradecahydro-2H-[1,3]dioxolo[4,5-c]1-oxa-6-azacyclopentadecan-15-yl]-8-oxooctyl}triphenylphosphoniumhexafluorophosphate(V)

A stirred solution of{8-[(2R,3S,4R,5R,8R,10R,11R,12S,13S,14R)-11-{[(2S,3R,4S,6R)-4-(dimethylamino)-3-hydroxy-6-methyloxan-2-yl]oxy}-2-ethyl-3,4,10-trihydroxy-13-{[(2R,4R,5S,6S)-5-hydroxy-4-methoxy-4,6-dimethyloxan-2-yl]oxy}-3,5,8,10,12,14-hexamethyl-15-oxo-1-oxa-6-azacyclopentadecan-6-yl]-8-oxooctyl}triphenylphosphoniumhexafluorophosphate(V) (prepared as described in Example 9) (55 mg, 0.04mmol) in DCM (10 mL) was treated with Ac₂O (8.2 μL, 0.09 mmol) and Et₃N(24 μL, 0.17 mmol). After stirring at room temperature for 16 h thereaction mixture was diluted with DCM (10 mL) and quenched withsaturated aqueous NaHCO₃ (10 mL) and H₂O (10 mL). The layers wereseparated and the aqueous further extracted with DCM (10 mL). Thecombined organic layers were dried over Na₂SO₄ and concentrated to givea white solid (60 mg), which was used without further purification inthe next step. A stirred solution of the white solid (60 mg, 0.05 mmol)in toluene (5 mL) and Et₃N (17 μL, 0.12 mmol) was treated withcarbonyldiimidazole (CDI) (59 mg, 0.37 mmol). The resulting reactionmixture was heated at 70° C. for 16 h. On cooling the reaction mixturewas diluted with EtOAc (15 mL) and the organic washed with saturatedaqueous NaHCO₃ (2×10 mL), The organics were dried over Na₂SO₄ andconcentrated under vacuo to give a further white solid (66 mg) which wasused without further purification in the next step. A stirred solutionof the white solid (66 mg, 0.05 mmol) in THF (5 mL) was treated with 0.1M NaOH (0.79 mL, 0.08 mmol) at 0 to 5° C. The reaction mixture was thenallowed to warm to room temperature. After stirring for a further 3 hthe reaction was diluted with saturated aqueous NH₄Cl (10 mL) andextracted with EtOAc (3×15 mL). The combined organics were concentratedunder reduced pressure and the resulting residue purified by columnchromatography eluting with 0-3% MeOH/0.7 M NH₃ in DCM to give{8-[(3aR,4R,7R,8S,9S,10R,11R,13R,16R,16aR)-10-{[(2S,3R,4S,6R)-3-(acetyloxy)-4-(dimethylamino)-6-methyloxan-2-yl]oxy}-4-ethyl-11-hydroxy-8-{[(2R,4R,5S,6S)-5-hydroxy-4-methoxy-4,6-dimethyloxan-2-yl]oxy}-3a,7,9,11,13,16-hexamethyl-2,6-dioxo-tetradecahydro-2H-[1,3]dioxolo[4,5-c]1-oxa-6-azacyclopentadecan-15-yl]-8-oxooctyl}triphenylphosphoniumhexafluorophosphate(V) as a colourless solid, which was taken up in MeOH(5 mL) and heated at 65° C. for 6 h. On allowing to cool the solvent wasremoved under reduced pressure to give the title compound (13 mg) as tancoloured solid.

LC-MS (Method H) 1148 [M]⁺; RT 7.26 min

Example11—(2S,3R,4S,6R)-4-(dimethylamino)-2-{[(2R,3R,4R,5R,8R,10R,11R,12S,13S,14R)-2-ethyl-3,10-dihydroxy-13-{[(2R,4R,5S,6S)-5-hydroxy-4-methoxy-4,6-dimethyloxan-2-yl]oxy}-3,5,6,8,10,12,14-heptamethyl-15-oxo-4-({[6-(triphenylphosphoniumyl)hexyl]carbamoyl}oxy)-1-oxa-6-azacyclopentadecan-11-yl]oxy}-6-methyloxan-3-ylacetate bromide

A stirred solution of(2S,3S,4R,6R)-6-{[(3aR,4R,7R,8S,9S,10R,11R,13R,16R,16aR)-10-{[(2S,3R,4S,6R)-3-(acetyloxy)-4-(dimethylamino)-6-methyloxan-2-yl]oxy}-4-ethyl-11-hydroxy-3a,7,9,11,13,15,16-heptamethyl-2,6-dioxo-tetradecahydro-2H-[1,3]dioxolo[4,5-c]1-oxa-6-azacyclopentadecan-8-yl]oxy}-4-methoxy-2,4-dimethyloxan-3-yl1H imidazole-1-carboxylate (prepared as described in European Journal ofMedicinal Chemistry 40, 2011, 5196) (2.0 g, 2.19 mmol) in THF (180 mL)was treated with 0.1 M NaOH (37.3 mL, 3.73 mmol) at 0 to 5° C. Thereaction mixture was then allowed to warm to room temperature. Afterstirring for a further 3 h the reaction was diluted with saturatedaqueous NH₄Cl (100 mL) and extracted with EtOAc (3×150 mL). The combinedorganics were concentrated under reduced pressure and the resultingresidue purified by column chromatography eluting with 0-10% MeOH/0.7 MNH₃ in DCM to give a white solid (1.32 g) which was used in the nextstep. A stirred solution of the white solid (0.21 g) and(6-aminohexyl)triphenylphosphonium bromide (prepared as described inWO2016025725) (0.34 g, 0.77 mmol) in pyridine (10 mL) was treated withDBU (0.11 mL, 0.77 mmol). After stirring for 15 d the reaction mixturewas diluted with EtOAc (10 mL) and DCM (5 mL) and the organics washedwith saturated aqueous NH₄Cl (3×10 mL). The organics were dried overNa₂SO₄ and concentrated in vacuo to give a yellow oil which was purifiedby 10% MeOH/0.7 M NH₃ in DCM to give the title compound as a white solid(16 mg). LC-MS (Method E) 1179 [M]⁺; RT 1.97 min

Example12—{12-[(2R,3S,4R,5R,8R,10R,11R,12S,13S,14R)-11-{[(2S,3R,4S,6R)-4-(dimethylamino)-3-hydroxy-6-methyloxan-2-yl]oxy}-2-ethyl-3,4,10,13-tetrahydroxy-3,5,8,10,12,14-hexamethyl-15-oxo-1-oxa-6-azacyclopentadecan-6-yl]-12-oxododecyl}triphenylphosphoniumhexafluorophosphate(V)

A solution of{12-[(2R,3S,4R,5R,8R,10R,11R,12S,13S,14R)-11-{[(2S,3R,4S,6R)-4-(dimethylamino)-3-hydroxy-6-methyloxan-2-yl]oxy}-2-ethyl-3,4,10-trihydroxy-13-{[(2R,4R,5S,6S)-5-hydroxy-4-methoxy-4,6-dimethyloxan-2-yl]oxy}-3,5,8,10,12,14-hexamethyl-15-oxo-1-oxa-6-azacyclopentadecan-6-yl]-12-oxododecyl}triphenylphosphoniumhexafluorophosphate(V) (prepared as described in Example 5) (0.26 g,0.61 mmol) in MeOH (10 mL) was treated with 10 M HCl (0.4 mL). Theresulting reaction mixture was stirred at room temperature for 16 h andthen neutralised with saturated aqueous NaHCO₃. The mixture wasconcentrated under reduced pressure and diluted with DCM. The DCM wasseparated and washed with 2 M HCl. The combined aqueous acidic fractionswere basified to pH 10 with 2 M NaOH and extracted with DCM. Thecombined organics from the basic extraction were dried over Na₂SO₄ andconcentrated under reduced pressure. The resulting residue was purifiedby column chromatography eluting with 2-10% MeOH/0.7 M NH₃ in DCM togive the title compound as a white solid.

LC-MS (Method G) 1020 [M]⁺; RT 1.87 min

Example13—{12-[(2R,3S,4R,5R,8R,10R,11R,12S,13S,14R)-2-ethyl-3,4,10,11,13-pentahydroxy-3,5,8,10,12,14-hexamethyl-15-oxo-1-oxa-6-azacyclopentadecan-6-yl]-12-oxododecyl}triphenylphosphoniumhexafluorophosphate(V)

Prepared following amide coupling procedure A1 using(2R,3S,4R,5R,8R,10R,11R,12S,13S,14R)-2-ethyl-3,4,10,11,13-pentahydroxy-3,5,8,10,12,14-hexamethyl-15-oxo-1-oxa-6-azacyclopentadecan-15-one(prepared as described in WO201111636312) and(11-carboxyundecyl)triphenylphosphonium bromide. Title compound isolatedas a white solid.

LC-MS (Method G) 863 [M]⁺; RT 1.65 min

Example14—{10-[(2R,3S,4R,5R,8R,10R,11R,12S,13S,14R)-11-{[(2S,3R,4S,6R)-4-(dimethylamino)-3-hydroxy-6-methyloxan-2-yl]oxy}-2-ethyl-3,4,10-trihydroxy-13-{[(2R,4R,5S,6S)-5-hydroxy-4-methoxy-4,6-dimethyloxan-2-yl]oxy}-3,5,8,10,12,14-hexamethyl-15-oxo-1-oxa-6-azacyclopentadecan-6-yl]decyl}triphenylphosphoniumiodide

Prepared following the general reductive amination procedure using10-chlorodecanal in step 1 and triphenylphosphine in step 2 underthermal heating conditions. Title compound isolated as a white solid.

1H NMR (Method A) (DMSO-d₆): δ (delta) ppm 7.94-7.87 (m, 3H), 7.84-7.74(m, 12H), 4.88 (dd, J=9.1 Hz, 1H), 4.80 (d, J=4.8 Hz, 1H), 4.44 (d,J=7.3 Hz, 1H), 4.32 (s, 1H), 4.22 (d, J=7.4 Hz, 1H), 4.12-3.98 (m, 2H),3.95-3.86 (m, 1H), 3.74-3.63 (m, 1H), 3.61-3.46 (m, 4H), 3.30 (s, 1H),3.23 (s, 3H), 3.14-2.98 (m, 1H), 2.95-2.88 (m, 1H), 2.86-2.75 (m, 1H),2.75-2.65 (m, 2H), 2.60-2.54 (m, 1H), 2.47-2.36 (m, 3H), 2.36-2.12 (m,6H), 2.12-2.01 (m, 1H), 2.00-1.83 (m, 2H), 1.82-1.70 (m, 1H), 1.68-1.32(m, 11H), 1.31-1.04 (m, 27H), 1.03-0.91 (m, 9H), 0.86 (d, J=6.6 Hz, 3H),0.80 (t, J=7.4 Hz, 3H); 31P NMR (162 MHz, DMSO-d₆): δ ppm+24.07 (s);LC-MS (Method H) 1136 [M]⁺; RT 9.10 min

Example15—{9-[(2R,3S,4R,5R,8R,10R,11R,12S,13S,14R)-11-{[(2S,3R,4S,6R)-4-(dimethylamino)-3-hydroxy-6-methyloxan-2-yl]oxy}-2-ethyl-3,4,10-trihydroxy-13-{[(2R,4R,5S,6S)-5-hydroxy-4-methoxy-4,6-dimethyloxan-2-yl]oxy}-3,5,8,10,12,14-hexamethyl-15-oxo-1-oxa-6-azacyclopentadecan-6-yl]nonyl}triphenylphosphoniumiodide

Prepared following the general reductive amination procedure using9-chlorononanal in step 1 and triphenylphosphine in step 2 under thermalheating conditions. Title compound isolated as a white solid.

LC-MS (Method H) 1122 [M]⁺; RT8.94 min

Example16—{14-[(2R,3S,4R,5R,8R,10R,11R,12S,13S,14R)-2-ethyl-3,4,10,11,13-pentahydroxy-3,5,8,10,12,14-hexamethyl-15-oxo-1-oxa-6-azacyclopentadecan-6-yl]-14-oxotetradecyl}triphenylphosphoniumhexafluorophosphate(V)

Prepared following amide coupling procedure A1 using(2R,3S,4R,5R,8R,10R,11R,12S,13S,14R)-2-ethyl-3,4,10,11,13-pentahydroxy-3,5,8,10,12,14-hexamethyl-15-oxo-1-oxa-6-azacyclopentadecan-15-one(prepared as described in WO201111636312) and(13-carboxytridecyl)triphenylphosphonium bromide. Title compoundisolated as an off white solid.

1H NMR (Method A) (DMSO-d₆): δ (delta) ppm 7.96-7.87 (m, 3H), 7.85-7.71(m, 12H), 5.14 (d, J=9.7 Hz, 1H), 4.70-4.60 (m, 2H), 4.52-4.44 (m, 1H),4.31-4.26 (m, 1H), 4.21-4.15 (m, 1H), 3.62-3.50 (m, 2H), 3.44-3.36 (m,1H), 3.27-3.21 (m, 1H), 3.21-3.16 (m, 1H), 3.05-2.96 (m, 1H), 2.42-2.29(m, 2H), 2.19-2.00 (m, 2H), 1.87-1.72 (m, 1H), 1.59-1.32 (m, 9H),1.31-1.10 (m, 25H), 1.10-1.00 (m, 4H), 0.91 (d, J=6.3 Hz, 3H), 0.88-0.82(m, 2H), 0.81-0.74 (m, 3H), 0.71-0.62 (m, 3H); 31P NMR (162 MHz,DMSO-d₆): δ ppm+24.05 (s);

LC-MS (Method E) 890.8 [M]⁺; RT 1.79 min

Example17—(2R,3R,4R,5R,8R,10R,11R,12S,13S,14R)-11-{[(2S,3R,4S,6R)-4-(dimethylamino)-3-hydroxy-6-methyloxan-2-yl]oxy}-2-ethyl-3,10-dihydroxy-13-{[(2R,4R,5S,6S)-5-hydroxy-4-methoxy-4,6-dimethyloxan-2-yl]oxy}-3,5,6,8,10,12,14-heptamethyl-15-oxo-1-oxa-6-azacyclopentadecan-4-ylN-[6-(triphenylphosphoniumyl)hexyl]carbamate bromide

A stirred solution of(2S,3R,4S,6R)-4-(dimethylamino)-2-{[(2R,3R,4R,5R,8R,10R,11R,12S,13S,14R)-2-ethyl-3,10-dihydroxy-13-{[(2R,4R,5S,6S)-5-hydroxy-4-methoxy-4,6-dimethyloxan-2-yl]oxy}-3,5,6,8,10,12,14-heptamethyl-15-oxo-4-({[6-(triphenylphosphosphoniumyl)hexyl]carbamoyl}oxy)-1-oxa-6-azacyclopentadecan-11-yl]oxy}-6-methyloxan-3-ylacetate bromide (prepared as described in Example 11) (41 mg, 0.03 mmol)in MeOH (5 mL) was stirred at room temperature. After 16 h the solventwas removed under reduced pressure to give the title compound (20 mg) asa white solid.

LC-MS (Method H) 1136.9 [M]⁺; RT 7.24 min

Example18—(2S,3R,4S,6R)-4-(dimethylamino)-2-{[(2R,3R,4R,5R,8R,10R,11R,12S,13S,14R)-2-ethyl-3,10-dihydroxy-13-{[(2R,4R,5S,6S)-5-hydroxy-4-methoxy-4,6-dimethyloxan-2-yl]oxy}-3,5,6,8,10,12,14-heptamethyl-15-oxo-4-({[10-(triphenylphosphosphoniumyl)decyl]carbamoyl}oxy)-1-oxa-6-azacyclopentadecan-11-yl]oxy}-6-methyloxan-3-ylacetate bromide[10-(1,3-dioxo-2,3-dihydro-1H-isoindol-2-yl)decyl]triphenylphosphoniumbromide

A mixture of 2-(10-bromodecyl)isoindoline-1,3-dione (1.43 g, 5.46 mmol)and triphenylphosphine (2 g, 5.46 mmol) in MeCN (15.6 mL) was heated at70° C. overnight. On cooling the solvent was removed in vacuo and theresulting residue purified by column chromatography eluting with 0-10%MeOH in DCM to give[10-(1,3-dioxo-2,3-dihydro-1H-isoindol-2-yl)decyl]triphenylphosphoniumbromide (1 g, 29%).

LC-MS (Method B) 548.4 [M]⁺; RT 2.06 min

(10-aminodecyl)triphenylphosphonium bromide

To a solution of[10-(1,3-dioxo-2,3-dihydro-1H-isoindol-2-yl)decyl]triphenylphosphoniumbromide (prepared as described in Example 18 step (a)) (0.8 g, 1.27mmol) in EtOH (10 mL) was added hydrazine hydrate (1.53 mL). Theresulting reaction mixture was refluxed for 16 h. On cooling the solventwas removed under vacuo and the resulting residue purified by columnchromatography eluting with 0-20% MeOH/5% NH₃ in DCM to give(10-aminodecyl)triphenylphosphonium bromide (0.38 g, 61%) as a whitesolid.

LC-MS (Method F) 418.4 [M]⁺; RT 1.52 min

(2S,3R,4S,6R)-4-(dimethylamino)-2-{[(2R,3R,4R,5R,8R,10R,11R,12S,13S,14R)-2-ethyl-3,10-dihydroxy-13-{[(2R,4R,5S,6S)-5-hydroxy-4-methoxy-4,6-dimethyloxan-2-yl]oxy}-3,5,6,8,10,12,14-heptamethyl-15-oxo-4-({[10-(triphenylphosphosphoniumyl)decyl]carbamoyl}oxy)-1-oxa-6-azacyclopentadecan-11-yl]oxy}-6-methyloxan-3-ylacetate bromide

Prepared following the procedure of Example 11 using(10-aminodecyl)triphenylphosphonium bromide (prepared as described inExample 18 step (b)). The title compound was isolated as a white solid.

1H NMR (Method A) (DMSO-d₆): δ (delta) ppm 7.94-7.85 (m, 3H), 7.84-7.72(m, 12H), 6.88 (t, 1H), 4.88 (dd, J=9.7 Hz, 1H), 4.73 (d, J=4.5 Hz, 1H),4.64-4.58 (m, 1H), 4.57-4.49 (m, 1H), 4.43-4.35 (m, 1H), 4.15-4.10 (m,1H), 4.07-3.99 (m, 1H), 3.99-3.93 (m, 1H), 3.72-3.61 (m, 1H), 3.60-3.49(m, 2H), 3.48-3.42 (m, 1H), 3.32 (s, 2H), 3.22 (s, 3H), 3.02-2.86 (m,4H), 2.77-2.70 (m, 1H), 2.70-2.59 (m, 1H), 2.44-2.35 (m, 1H), 2.28 (d,J=14.9 Hz, 1H), 2.15 (s, 6H), 2.06 (s, 3H), 1.93 (s, 4H), 1.90-1.66 (m,5H), 1.64-1.30 (m, 8H), 1.28-1.03 (m, 29H), 0.98 (d, J=9.2 Hz, 1H),0.92-0.81 (m, 9H), 0.76 (t, J=7.3 Hz, 3H); 31P NMR (162 MHz, DMSO-d₆): δppm+24.04 (s); LC-MS (Method H) 1235 [M]⁺; RT 9.32 min

Example19—(2R,3R,4R,5R,8R,10R,11R,12S,13S,14R)-11-{[(2S,3R,4S,6R)-4-(dimethylamino)-3-hydroxy-6-methyloxan-2-yl]oxy}-2-ethyl-3,10-dihydroxy-13-{[(2R,4R,5S,6S)-5-hydroxy-4-methoxy-4,6-dimethyloxan-2-yl]oxy}-3,5,6,8,10,12,14-heptamethyl-15-oxo-1-oxa-6-azacyclopentadecan-4-ylN-[10-(triphenylphosphoniumyl)decyl]carbamate bromide

Prepared following the procedure of Example 17 using(2S,3R,4S,6R)-4-(dimethylamino)-2-{[(2R,3R,4R,5R,8R,10R,11R,12S,13S,14R)-2-ethyl-3,10-dihydroxy-13-{[(2R,4R,5S,6S)-5-hydroxy-4-methoxy-4,6-dimethyloxan-2-yl]oxy}-3,5,6,8,10,12,14-heptamethyl-15-oxo-4-({[10-(triphenylphosphosphoniumyl)decyl]carbamoyl}oxy)-1-oxa-6-azacyclopentadecan-11-yl]oxy}-6-methyloxan-3-ylacetate bromide (prepared as described in Example 18). Purification wasby chromatography eluting with 0-10% MeOH/1% NH₃ in DCM to give thetitle compound as a white solid.

1H NMR (Method A) (DMSO-d₆): δ (delta) ppm 7.96-7.85 (m, 3H), 7.80 (dtd,J=10.8, 8.3, 2.9 Hz, 12H), 6.86 (t, J=6.4 Hz, 1H), 4.98 (d, J=9.3 Hz,1H), 4.73 (d, J=4.6 Hz, 1H), 4.60 (d, J=7.2 Hz, 1H), 4.40 (s, 1H), 4.36(d, J=7.1 Hz, 1H), 4.28 (d, J=8.5 Hz, 1H), 4.18 (d, J=8.0 Hz, 1H),4.12-3.98 (m, 1H), 3.81 (s, 1H), 3.63-3.50 (m, 3H), 3.45 (d, J=7.3 Hz,1H), 3.22-3.17 (m, 3H), 3.05 (d, J=7.6 Hz, 1H), 3.01-2.93 (m, 1H),2.93-2.84 (m, 2H), 2.73-2.64 (m, 1H), 2.43-2.32 (m, 1H), 2.24 (s, 7H),2.07 (s, 3H), 2.03-1.95 (m, 1H), 1.92-1.68 (m, 4H), 1.67-1.58 (m, 2H),1.56-1.32 (m, 8H), 1.30-0.98 (m, 35H), 0.90 (d, J=6.4 Hz, 3H), 0.85 (d,J=6.4 Hz, 3H), 0.80 (t, J=7.4 Hz, 3H); 31P NMR (162 MHz, DMSO-d₆): δppm+24.07 (s); LC-MS (Method H) 1193[M]⁺; RT 8.65 min

Example20-(2R,3R,4R,5R,8R,10R,11R,12S,13S,14R)-11-{[(2S,3R,4S,6R)-4-(dimethylamino)-3-hydroxy-6-methyloxan-2-yl]oxy}-2-ethyl-3,10-dihydroxy-13-{[(2R,4R,5S,6S)-5-hydroxy-4-methoxy-4,6-dimethyloxan-2-yl]oxy}-3,5,6,8,10,12,14-heptamethyl-15-oxo-1-oxa-6-azacyclopentadecan-4-ylN-[8-(triphenylphosphoniumyl)octyl]carbamate bromide8-(1,3-dioxo-2,3-dihydro-1H-isoindol-2-yl)octyl]triphenylphosphoniumbromide

Prepared following the procedure of Example 18 step (a) using2-(8-bromooctyl)isoindole-1,3-dione.

LC-MS (Method B) 520.3 [M]⁺; RT 1.85 min

(8-aminooctyl)triphenylphosphonium bromide

Prepared following the procedure of Example 18 step (b) using[8-(1,3-dioxo-2,3-dihydro-1H-isoindol-2-yl)octyl]triphenylphosphoniumbromide (prepared as described in Example step (a)). LC-MS (Method B)390.3 [M]⁺; RT 0.72 min

(2S,3R,4S,6R)-4-(dimethylamino)-2-{[(2R,3R,4R,5R,8R,10R,11R,12S,13S,14R)-2-ethyl-3,10-dihydroxy-13-{[(2R,4R,5S,6S)-5-hydroxy-4-methoxy-4,6-dimethyloxan-2-yl]oxy}-3,5,6,8,10,12,14-heptamethyl-15-oxo-4-({[8-(triphenylphosphosphoniumyl)octyl]carbamoyl}oxy)-1-oxa-6-azacyclopentadecan-11-yl]oxy}-6-methyloxan-3-ylacetate bromide

Prepared following the procedure of Example 11 using(8-aminooctyl)triphenylphosphonium bromide (prepared as described inExample 20 step (b)), and used in the next step.

LC-MS (Method H) 1206.8 [M]⁺; RT 9.03 min

(2R,3R,4R,5R,8R,10R,11R,12S,13S,14R)-11-{[(2S,3R,4S,6R)-4-(dimethylamino)-3-hydroxy-6-methyloxan-2-yl]oxy}-2-ethyl-3,10-dihydroxy-13-{[(2R,4R,5S,6S)-5-hydroxy-4-methoxy-4,6-dimethyloxan-2-yl]oxy}-3,5,6,8,10,12,14-heptamethyl-15-oxo-1-oxa-6-azacyclopentadecan-4-ylN-[8-(triphenylphosphoniumyl)octyl]carbamate bromide

Prepared following the procedure of Example 17 using(2S,3R,4S,6R)-4-(dimethylamino)-2-{[(2R,3R,4R,5R,8R,10R,11R,12S,13S,14R)-2-ethyl-3,10-dihydroxy-13-{[(2R,4R,5S,6S)-5-hydroxy-4-methoxy-4,6-dimethyloxan-2-yl]oxy}-3,5,6,8,10,12,14-heptamethyl-15-oxo-4-({[8-(triphenylphosphosphoniumyl)octyl]carbamoyl}oxy)-1-oxa-6-azacyclopentadecan-11-yl]oxy}-6-methyloxan-3-ylacetate bromide (prepared as described in Example 20 step (c)).Purification was by chromatography eluting with 0-10% MeOH/1% NH₃ in DCMto give the title compound as a white solid.

LC-MS (Method H) 1164.8 [M]⁺; RT8.14 min

Example21—{14-[(2R,3S,4R,5R,8R,10R,11R,12S,13S,14R)-11-{[(2S,3R,4S,6R)-4-(dimethylamino)-3-hydroxy-6-methyloxan-2-yl]oxy}-2-ethyl-3,4,10,13-tetrahydroxy-3,5,8,10,12,14-hexamethyl-15-oxo-1-oxa-6-azacyclopentadecan-6-yl]-14-oxotetradecyl}triphenylphosphoniumhexafluorophosphate(V)

Prepared following the procedure in Example 12 using{14-[(2R,3S,4R,5R,8R,10R,11R,12S,13S,14R)-11-{[(2S,3R,4S,6R)-4-(dimethylamino)-3-hydroxy-6-methyloxan-2-yl]oxy}-2-ethyl-3,4,10-trihydroxy-13-{[(2R,4R,5S,6S)-5-hydroxy-4-methoxy-4,6-dimethyloxan-2-yl]oxy}-3,5,8,10,12,14-hexamethyl-15-oxo-1-oxa-6-azacyclopentadecan-6-yl]-14-oxotetradecyl}triphenylphosphoniumhexafluorophosphate(V) (prepared as described in Example 1).Purification was by chromatography eluting with 0-5% MeOH/0.7 M NH₃ inDCM to give the title compound as a white solid.

LC-MS (Method G) 1048 [M]⁺; RT 2.07 min

Example22—(2-{6-[(2R,3S,4R,5R,8R,10R,11R,12S,13S,14R)-11-{[(2S,3R,4S,6R)-4-(dimethylamino)-3-hydroxy-6-methyloxan-2-yl]oxy}-2-ethyl-3,4,10-trihydroxy-13-{[(2R,4R,5S,6S)-5-hydroxy-4-methoxy-4,6-dimethyloxan-2-yl]oxy}-3,5,6,8,10,12,14-heptamethyl-15-oxo-1-oxa-6-azacyclopentadecan-4-ylN-methylhexanamido}ethyl)triphenylphosphonium bromide benzyl6-[(2R,3S,4R,5R,8R,10R,11R,12S,13S,14R)-11-{[(2S,3R,4S,6R-4-(dimethylamino)-3-hydroxy-6-methyloxan-2-yl]oxy}-2-ethyl-3,4,10-trihydroxy-13-{[(2R,4R,5S,6S)-5-hydroxy-4-methoxy-4,6-dimethyloxan-2-yl]oxy}-3,5,6,8,10,12,14-hexamethyl-15-oxo-1-oxa-6-azacyclopentadecan-6-yl]hexanoate

To a stirred solution of N-desmethyl-azithromycin (0.85 g, 1.16 mmol),benzyl 6-oxohexanoate (0.64 g, 2.91 mmol) and CH₃COOH (0.67 mL, 11.6mmol) in DMF (30 mL) was added NaBH₃CN (0.15 g, 2.32 mmol). Theresulting mixture was heated at 70° C. for 16 h. On cooling the reactionwas quenched with H₂O (20 mL), saturated aqueous NaHCO₃ (10 mL) anddiluted with DCM (30 mL). The organic layer was separated and theaqueous further extracted with DCM (2×30 mL). The combined organics werewashed with saturated aqueous NaHCO₃ (50 mL) and brine (50 mL). Afterdrying over Na₂SO₄ the organics were concentrated under reduced pressureand the resulting residue purified by chromatography eluting with 5-7%MeOH/0.7 M NH₃ in DCM to give benzyl6-[(2R,3S,4R,5R,8R,10R,11R,12S,13S,14R)-11-{[(2S,3R,4S,6R)-4-(dimethylamino)-3-hydroxy-6-methyloxan-2-yl]oxy}-2-ethyl-3,4,10-trihydroxy-13-{[(2R,4R,5S,6S)-5-hydroxy-4-methoxy-4,6-dimethyloxan-2-yl]oxy}-3,5,6,8,10,12,14-hexamethyl-15-oxo-1-oxa-6-azacyclopentadecan-6-yl]hexanoate(0.45 g) as a white solid, which was used in the next step.

6-[(2R,3S,4R,5R,8R,10R,11R,12S,13S,14R)-11-{[(2S,3R,4S,6R)-4-(dimethylamino)-3-hydroxy-6-methyloxan-2-yl]oxy}-2-ethyl-3,4,10-trihydroxy-13-{[(2R,4R,5S,6S)-5-hydroxy-4-methoxy-4,6-dimethyloxan-2-yl]oxy}-3,5,6,8,10,12,14-hexamethyl-15-oxo-1-oxa-6-azacyclopentadecan-6-yl]hexanoicacid

Pd/C (50 mg) was added to a solution of benzyl6-[(2R,3S,4R,5R,8R,10R,11R,12S,13S,14R)-11-{[(2S,3R,4S,6R)-4-(dimethylamino)-3-hydroxy-6-methyloxan-2-yl]oxy}-2-ethyl-3,4,10-trihydroxy-13-{[(2R,4R,5S,6S)-5-hydroxy-4-methoxy-4,6-dimethyloxan-2-yl]oxy}-3,5,6,8,10,12,14-hexamethyl-15-oxo-1-oxa-6-azacyclopentadecan-6-yl]hexanoate(prepared as described in Example 22 step (a)) (0.45 g, 0.47 mmol) inEtOH (20 mL) and CH₃COOH (2 drops). The reaction mixture was placed in ahydrogenation autoclave and purged with N₂ (3×) followed by H₂ (3×). Thereaction mixture was then stirred at room temperature under H₂ at 5 bar.After 16 h the reaction mixture was filtered through a pad of celite,which was washed with EtOH (20 mL). The organics were concentrated underreduced pressure and the resulting residue purified by chromatographyeluting with 0-20% MeOH/0.7 M NH₃ in DCM to give6-[(2R,3S,4R,5R,8R,10R,11R,12S,13S,14R)-11-{[(2S,3R,4S,6R)-4-(dimethylamino)-3-hydroxy-6-methyloxan-2-yl]oxy}-2-ethyl-3,4,10-trihydroxy-13-{[(2R,4R,5S,6S)-5-hydroxy-4-methoxy-4,6-dimethyloxan-2-yl]oxy}-3,5,6,8,10,12,14-hexamethyl-15-oxo-1-oxa-6-azacyclopentadecan-6-yl]hexanoicacid (0.20 g) as a white solid, which was used in the next step.

LC-MS (Method H) 850 [M+H]⁺; RT 3.74 min

(2-{6-[(2R,3S,4R,5R,8R,10R,11R,12S,13S,14R)-11-{[(2S,3R,4S,6R)-4-(dimethylamino)-3-hydroxy-6-methyloxan-2-yl]oxy}-2-ethyl-3,4,10-trihydroxy-13-{[(2R,4R,5S,6S)-5-hydroxy-4-methoxy-4,6-dimethyloxan-2-yl]oxy}-3,5,6,8,10,12,14-heptamethyl-15-oxo-1-oxa-6-azacyclopentadecan-4-ylN-methylhexanamido}ethyl)triphenylphosphonium bromide

To a solution of6-[(2R,3S,4R,5R,8R,10R,11R,12S,13S,14R)-11-{[(2S,3R,4S,6R)-4-(dimethylamino)-3-hydroxy-6-methyloxan-2-yl]oxy}-2-ethyl-3,4,10-trihydroxy-13-{[(2R,4R,5S,6S)-5-hydroxy-4-methoxy-4,6-dimethyloxan-2-yl]oxy}-3,5,6,8,10,12,14-hexamethyl-15-oxo-1-oxa-6-azacyclopentadecan-6-yl]hexanoicacid (prepared as described in Example 22 step (b)) (75 mg, 0.09 mmol),[2-(methylamino)ethyl]triphenylphosphonium bromide (42 mg, 0.13 mmol)and 1-hydroxy-7-azabenzotriazole (HOAt) (15 mg, 0.1 mmol) in DCM (10 mL)was added N,N′-diisopropylcarbodiimide (DIC) (15 μL, 0.10 mmol). Theresulting reaction mixture was heated at 30° C. for 16 h. On cooling thereaction mixture was diluted with DCM (10 mL) and washed with saturatedaqueous NaHCO₃ (5 mL) and H₂O (10 mL). The combined aqueous extractswere back extracted with DCM (2×10 mL). The combined organic extractswere dried over Na₂SO₄ and concentrated under reduced pressure. Theresulting residue was purified by chromatography eluting with 0-10%MeOH/0.7 M NH₃ in DCM to give the title compound (56 mg) as a colourlesssolid.

1H NMR (Method A) (DMSO-d₆): δ (delta) ppm 7.93-7.81 (m, 9H), 7.81-7.74(m, 6H), 4.89 (dd, J=10.0 Hz, 1H), 4.80 (d, J=4.8 Hz, 1H), 4.44 (d,J=7.2 Hz, 1H), 4.33 (s, 1H), 4.23 (d, J=7.5 Hz, 1H), 4.10-3.98 (m, 3H),3.98-3.91 (m, 1H), 3.85-3.76 (m, 2H), 3.73-3.56 (m, 3H), 3.56-3.47 (m,2H), 3.23 (s, 3H), 3.12-3.04 (m, 1H), 2.96 (s, 3H), 2.91 (t, J=8.4 Hz,1H), 2.83 (s, 1H), 2.76-2.69 (m, 2H), 2.62-2.54 (m, 1H), 2.46-2.18 (m,8H), 2.15-2.08 (m, 3H), 2.07-1.84 (m, 3H), 1.77 (d, J=6.3 Hz, 2H),1.57-1.47 (m, 2H), 1.46-1.28 (m, 6H), 1.27-1.05 (m, 19H), 1.05-0.93 (m,9H), 0.87 (d, J=6.6 Hz, 3H), 0.80 (t, J=7.4 Hz, 3H); 31P NMR (162 MHz,DMSO-d₆): δ ppm+21.42 (s); LC-MS (Method H) 1151 [M]⁺; RT 7.32 min

Example23—[2-(2-{3-[(2R,3S,4R,5R,8R,10R,11R,12S,13S,14R)-11-{[(2S,3R,4S,6R)-4-(dimethylamino)-3-hydroxy-6-methyloxan-2-yl]oxy}-2-ethyl-3,4,10-trihydroxy-13-{[(2R,4R,5S,6S)-5-hydroxy-4-methoxy-4,6-dimethyloxan-2-yl]oxy}-3,5,6,8,10,12,14-hexamethyl-15-oxo-1-oxa-6-azacyclopentadecan-6-yl]-3-oxopropoxy}ethoxy)ethyl]triphenylphosphoniumbromide(2-{2-[3-(tert-butoxy)-3-oxopropoxy]ethoxy}ethyl)triphenylphosphoniumbromide

To a solution of triphenyl(vinyl)phosphonium bromide (1.62 g, 4.38 mmol)and tert-butyl 3-(2-hydroxyethoxy)propanoate (1.24 g, 6 mmol) in dry THF(15 mL) was added 2.5 M butyllithium (0.18 mL, 0.44 mmol). The resultingreaction was stirred at room temperature. After 24 h the reactionmixture was quenched with MeOH (5 mL) and concentrated under reducedpressure. The resulting residue was purified by chromatography elutingwith 0-10% MeOH/0.7 M NH₃ in DCM to give(2-{2-[3-(tert-butoxy)-3-oxopropoxy]ethoxy}ethyl)triphenylphosphoniumbromide (0.92 g) as a sticky opague oil, which was used in the next stepwithout any further purification.

{2-[2-(2-carboxyethoxy)ethoxy]ethyl}triphenylphosphonium bromide

A stirred solution of(2-{2-[3-(tert-butoxy)-3-oxopropoxy]ethoxy}ethyl)triphenylphosphoniumbromide (prepared as described in Example 23 step (a)) (0.92 g, 1.65mmol) in DCM (10 mL) at 35° C. was treated with 4 M HCl in dioxan (2.06mL). After 16 h the reaction mixture was allowed cool and concentratedunder reduced pressure co-evaporating with MeCN (2×10 mL) to give{2-[2-(2-carboxyethoxy)ethoxy]ethyl}triphenylphosphonium bromide as awhite solid, which was used without further purification in the nextstep.

[2-(2-{3-[(2R,3S,4R,5R,8R,10R,11R,12S,13S,14R)-11-{[(2S,3R,4S,6R)-4-(dimethylamino)-3-hydroxy-6-methyloxan-2-yl]oxy}-2-ethyl-3,4,10-trihydroxy-13-{[(2R,4R,5S,6S)-5-hydroxy-4-methoxy-4,6-dimethyloxan-2-yl]oxy}-3,5,6,8,10,12,14-hexamethyl-15-oxo-1-oxa-6-azacyclopentadecan-6-yl]-3-oxopropoxy}ethoxy)ethyl]triphenylphosphoniumbromide

To a solution of N-desmethyl-azithromycin (0.2 g, 0.27 mmol),{2-[2-(2-carboxyethoxy)ethoxy]ethyl}triphenylphosphonium bromide(prepared as described in Example 23 step (b)) (0.27 g, 0.54 mmol) and1-hydroxy-7-azabenzotriazole (HOAt) (46 mg, 0.3 mmol) in DCM (10 mL) wasadded N,N′-diisopropylcarbodiimide (DIC) (47 μL, 0.3 mmol). Theresulting reaction mixture was heated at 30° C. After 16 h furtherquantities of {2-[2-(2-carboxyethoxy)ethoxy]ethyl}triphenylphosphoniumbromide (0.13 g, 0.27 mmol) and N,N′-diisopropylcarbodiimide (DIC) (47μL, 0.3 mmol) were added and heating continued for a further 16 h. Oncooling the reaction mixture was concentrated under reduced pressure andthe resulting residue taken up in MeOH (20 mL) and heated at 50° C. for16 h. On cooling the solvent was removed under reduced pressure and theresulting residue purified by chromatography eluting with 0-20% MeOH/0.7M NH₃ in DCM to give the title compound (15 mg) as a white solid.

LC-MS (Method H) 1151 [M]⁺; RT 6.66 min

Example24-[2-({5-[(2R,3S,4R,5R,8R,10R,11R,12S,13S,14R)-11-{[(2S,3R,4S,6R)-4-(dimethylamino)-3-hydroxy-6-methyloxan-2-yl]oxy}-2-ethyl-3,4,10-trihydroxy-13-{[(2R,4R,5S,6S)-5-hydroxy-4-methoxy-4,6-dimethyloxan-2-yl]oxy}-3,5,6,8,10,12,14-hexamethyl-15-oxo-1-oxa-6-azacyclopentadecan-6-yl]-5-oxopentyl}(methyl)amino)ethyl]triphenylphosphoniumbromide(2-{[5-(tert-butoxy)-5-oxopentyl](methyl)amino}ethyl)triphenylphosphoniumbromide

To a stirred solution of (2-(methylamino)ethyl)triphenylphosphoniumbromide (1.24 g, 3.11 mmol) and N,N-diisopropylethyleneamine (DIEA)(0.65 mL, 3.73 mmol) in MeCN was added tert-butyl 5-bromopentanoate (0.7g, 2.95 mmol). The resulting reaction mixture was heated at 50° C. for16 h. On cooling the solvent was removed under reduced pressure and theresidue taken up in 10% MeOH in DCM (30 mL). The organics were washedwith H₂O (10 mL) and saturated aqueous NaHCO₃ (10 mL). The combinedaqueous extracts were back extracted with 10% MeOH in DCM (3×30 mL). Thecombined organic extracts were dried over Na₂SO₄ and concentrated underreduced pressure. The resulting residue was purified by chromatographyeluting with 0-10% MeOH/0.7 M NH₃ in DCM to give(2-{[5-(tert-butoxy)-5-oxopentyl](methyl)amino}ethyl)triphenylphosphoniumbromide (0.74 g) as a clear oil, which was used in the next step.

{2-[(4-carboxybutyl)(methyl)amino]ethyl}triphenylphosphonium bromide

Prepared following the procedure in Example 23 step (b) using(2-{[5-(tert-butoxy)-5-oxopentyl](methyl)amino}ethyl)triphenylphosphoniumbromide (prepared as described in Example 24 step (a)). The resulting{2-[(4-carboxybutyl)(methyl)amino]ethyl}triphenylphosphonium bromide wasisolated as a white solid, which on standing turned to a yellow oil, andwas used in the next step without any further purification.

[2-({5-[(2R,3S,4R,5R,8R,10R,11R,12S,13S,14R)-11-{[(2S,3R,4S,6R)-4-(dimethylamino)-3-hydroxy-6-methyloxan-2-yl]oxy}-2-ethyl-3,4,10-trihydroxy-13-{[(2R,4R,5S,6S)-5-hydroxy-4-methoxy-4,6-dimethyloxan-2-yl]oxy}-3,5,6,8,10,12,14-hexamethyl-15-oxo-1-oxa-6-azacyclopentadecan-6-yl]-5-oxopentyl}(methyl)amino)ethyl]triphenylphosphoniumbromide

Prepared following the procedure in Example 23 step (c) using{2-[(4-carboxybutyl)(methyl)amino]ethyl}triphenylphosphonium bromide(prepared as described in Example 24 step (b)). Purification was bychromatography eluting with 0-30% MeOH/0.7 M NH₃ in DCM to give thetitle compound as a white solid.

LC-MS (Method H) 1137 [M]⁺; RT 7.11 min

Example25—{11-[(2R,3S,4R,5R,8R,10R,11R,12S,13S,14R)-11-{[(2S,3R,4S,6R)-4-(dimethylamino)-3-hydroxy-6-methyloxan-2-yl]oxy}-2-ethyl-3,4,10-trihydroxy-13-{[(2R,4R,5S,6S)-5-hydroxy-4-methoxy-4,6-dimethyloxan-2-yl]oxy}-3,5,8,10,12,14-hexamethyl-15-oxo-1-oxa-6-azacyclopentadecan-6-yl]-11-oxoundecyl}triphenylphosphoniumchloride

A stirred solution of (10-carboxydecyl)triphenylphosphonium bromide (431mg, 0.82 mmol), 1-hydroxy-7-azabenzotriazole (HOAt) (111 mg, 0.82 mmol),N,N-diisopropylethylamine (DIEA) (0.36 mL, 2.04 mmol) andN,N′-diisopropylcarbodiimide (DIC) (127 μL, 0.82 mmol) in DCM (2 mL) washeated at 40° C. After 30 min the reaction mixture was added to astirred solution of N-desmethyl-azithromycin (0.5 g. 0.68 mmol) in DCM(2 mL) at 40° C. Stirring at 40° C. was continued overnight. On coolingto room temperature H₂O (5 mL) and DCM (5 mL) were added and the mixturepassed through a phase separator, washing with DCM (2×1 mL). The DCMfractions wee pooled and solvent removed under reduced pressure to givea residue, which was taken up in MeOH (5 mL) and heated at 60° C. for 2h. On cooling the solvent was removed under reduced pressure and theresulting residue taken up in THF (10 mL), which was washed withsaturated aqueous NH₄Cl (10 mL). The separated aqueous phase was furtherextracted with THF (2×5 mL). The combined organic extracts were washedwith brine (10 mL), dried over MgSO₄ and solvent removed under reducedpressure. The resulting residue was purified by silica columnchromatography eluting with 0-10% MeOH/0.7 M NH₃ in DCM. The resultingproduct was partitioned between DCM (10 mL) and saturated aqueous NH₄Cl(10 mL). The separated organic phase was passed through a phaseseparator containing brine (5 mL). The collected solvent was evaporatedunder reduced pressure and the resulting residue taken up in MeOH (3 mL)and filtered through an Amberlite IRA-400(Cl) ion exchange resin. Thecollected MeOH was recycled through the column 3×, followed by a freshvolume of MeOH. The combined MeOH washings were concentrated underreduced pressure to give the title compound (0.4 g) as a white solid.

LC-MS (Method D) 1164 [M]⁺; RT 1.93 min

Example26—(2S,3R,4S,6R)-4-(dimethylamino)-2-{[(2R,3R,4R,5R,8R,10R,11R,12S,13S,14R)-2-ethyl-3,10,13-trihydroxy-3,5,6,8,10,12,14-heptamethyl-15-oxo-4-({[8-(triphenylphosphoniumyl)octyl]carbamoyl}oxy)-1-oxa-6-azacyclopentadecan-11-yl]oxy}-6-methyloxan-3-ylacetate bromide(2S,3R,4S,6R)-2-{[(3aR,4R,7R,8S,9S,10R,11R,13R,16R,16aR)-4-ethyl-11-hydroxy-8-{[(2R,4R,5S,6S)-4-methoxy-5-[(methoxycarbonyl)oxy]-4,6-dimethyloxan-2-yl]oxy}-3a,7,9,11,13,15,16-heptamethyl-2,6-dioxo-tetradecahydro-2H-[1,3]dioxolo[4,5-c]1-oxa-6-azacyclopentadecan-10-yl]oxy}-4-(dimethylamino)-6-methyloxan-3-ylacetate

A stirred solution of(2S,3S,4R,6R)-6-{[(3aR,4R,7R,8S,9S,10R,11R,13R,16R,16aR)-10-{[(2S,3R,4S,6R)-3-(acetyloxy)-4-(dimethylamino)-6-methyloxan-2-yl]oxy}-4-ethyl-11-hydroxy-3a,7,9,11,13,15,16-heptamethyl-2,6-dioxo-tetradecahydro-2H-[1,3]dioxolo[4,5-c]1-oxa-6-azacyclopentadecan-8-yl]oxy}-4-methoxy-2,4-dimethyloxan-3-yl1H imidazole-1-carboxylate (prepared as described in European Journal ofMedicinal Chemistry 40, 2011, 5196) (1.0 g, 1.1 mmol) in MeOH (53.3 mL)was treated with 12 M HCl (0.46 mL). The resulting reaction mixture wasstirred at room temperature. After 15 h further 12 M HCl (0.46 mL) wasadded and stirring continued. After a further 15 h the reaction mixturewas neutralised with solid NaHCO₃ and solvent removed to give a whitesolid, which was purified by silica column chromatography eluting with0-100% acetone in i-hexane to give(2S,3R,4S,6R)-2-{[(3aR,4R,7R,8S,9S,10R,11R,13R,16R,16aR)-4-ethyl-11-hydroxy-8-{[(2R,4R,5S,6S)-4-methoxy-5-[(methoxycarbonyl)oxy]-4,6-dimethyloxan-2-yl]oxy}-3a,7,9,11,13,15,16-heptamethyl-2,6-dioxo-tetradecahydro-2H-[1,3]dioxolo[4,5-c]1-oxa-6-azacyclopentadecan-10-yl]oxy}-4-(dimethylamino)-6-methyloxan-3-ylacetate (0.46 g) as a white solid, which was used in the next step LC-MS(Method E) 876 [M+1]⁺; RT 1.99 min(2S,3R,4S,6R)-2-{[(3aR,4R,7R,8S,9S,10R,11R,13R,16R,16aR)-4-ethyl-8,11-dihydroxy-3a,7,9,11,13,15,16-heptamethyl-2,6-dioxo-tetradecahydro-2H-[1,3]dioxolo[4,5-c]1-oxa-6-azacyclopentadecan-10-yl]oxy}-4-(dimethylamino)-6-methyloxan-3-ylacetate

A solution of(2S,3R,4S,6R)-2-{[(3aR,4R,7R,8S,9S,10R,11R,13R,16R,16aR)-4-ethyl-11-hydroxy-8-{[(2R,4R,5S,6S)-4-methoxy-5-[(methoxycarbonyl)oxy]-4,6-dimethyloxan-2-yl]oxy}-3a,7,9,11,13,15,16-heptamethyl-2,6-dioxo-tetradecahydro-2H-[1,3]dioxolo[4,5-c]1-oxa-6-azacyclopentadecan-10-yl]oxy}-4-(dimethylamino)-6-methyloxan-3-ylacetate (prepared as described in Example 26 step (a)) (0.46 g, 0.53mmol) in 0.25 M aqueous HCl was stirred at room temperature. After 6 dthe reaction mixture was washed with DCM (2×60 mL). The pH of theseparated aqueous was adjusted to approximately 10 using 2 N aqueousNaOH and extracted with DCM (2×60 mL). The combined organics were driedover Na₂SO₄ and solvent removed under reduced pressure. The resultingresidue was purified by silica column chromatography eluting initiallywith DCM followed by a solvent mixture of DCM/MeOH/NH₃ (90/10/1) to give(2S,3R,4S,6R)-2-{[(3aR,4R,7R,8S,9S,10R,11R,13R,16R,16aR)-4-ethyl-8,11-dihydroxy-3a,7,9,11,13,15,16-heptamethyl-2,6-dioxo-tetradecahydro-2H-[1,3]dioxolo[4,5-c]1-oxa-6-azacyclopentadecan-10-yl]oxy}-4-(dimethylamino)-6-methyloxan-3-ylacetate (0.13 g) as a white solid, which was used in the next step.LC-MS (Method E) 659 [M+1]⁺; RT 1.60 min

(2S,3R,4S,6R)-4-(dimethylamino)-2-{[(2R,3R,4R,5R,8R,10R,11R,12S,13S,14R)-2-ethyl-3,10,13-trihydroxy-3,5,6,8,10,12,14-heptamethyl-15-oxo-4-({[8-(triphenylphosphoniumyl)octyl]carbamoyl}oxy)-1-oxa-6-azacyclopentadecan-11-yl]oxy}-6-methyloxan-3-ylacetate bromide

Prepared following the procedure of Example 11 using(8-aminooctyl)triphenylphosphonium bromide (prepared as described inExample 20 step (b)) and(2S,3R,4S,6R)-2-{[(3aR,4R,7R,8S,9S,10R,11R,13R,16R,16aR)-4-ethyl-8,11-dihydroxy-3a,7,9,11,13,15,16-heptamethyl-2,6-dioxo-tetradecahydro-2H-[1,3]dioxolo[4,5-c]1-oxa-6-azacyclopentadecan-10-yl]oxy}-4-(dimethylamino)-6-methyloxan-3-ylacetate (prepared as described in Example 26 step (b)). Purification wasby silica column chromatography eluting initially with DCM followed by asolvent mixture of DCM/MeOH/NH₃ (90/10/1).

31P NMR (162 MHz, DMSO-d₆): δ ppm+24.06 (s); LC-MS (Method H) 1049 [M]⁺;RT 8.01 min

Example27—13-[(2R,3S,4R,5R,8R,10R,11R,12S,13S,14R)-11-{[(2S,3R,4S,6R)-4-(dimethylamino)-3-hydroxy-6-methyloxan-2-yl]oxy}-2-ethyl-3,4,10-trihydroxy-13-{[(2R,4R,5S,6S)-5-hydroxy-4-methoxy-4,6-dimethyloxan-2-yl]oxy}-3,5,8,10,12,14-hexamethyl-15-oxo-1-oxa-6-azacyclopentadecan-6-yl]-13-oxo-1,1,1-triphenyl-4,7,10-trioxa-1-phosphatridecan-1-iumchloride13-(tert-butoxy)-13-oxo-1,1,1-triphenyl-4,7,10-trioxa-1-phosphatridecan-1-iumiodide

A suspension of tert-butyl3-(2-{2-[2-(methanesulphonyloxy)ethoxy]ethoxy}ethoxy)propanoate (1.3 g,3.65 mmol), triphenylphosphine (0.96 g, 3.65 mmol) and NaI (0.55 g, 3.65mmol) in MeCN (10 mL) was degassed with bubbling N₂ for 15 min and thenheated at 80° C. under N₂ for 16 h. On cooling to room temperature, thesolution was filtered, and the solvent removed under reduced pressure.The resulting residue was purified by silica column chromatographyeluting with 0-10% MeOH/0.7 M NH₃ in DCM to give13-(tert-butoxy)-13-oxo-1,1,1-triphenyl-4,7,10-trioxa-1-phosphatridecan-1-iumiodide (2 q) as a yellow oil, which was used in the next step. LC-MS(Method I) 523 [M]⁺; RT 1.13 min

12-carboxy-1,1,1-triphenyl-4,7,10-trioxa-1-phospha dodecan-1-ium iodide

A stirred solution of13-(tert-butoxy)-13-oxo-1,1,1-triphenyl-4,7,10-trioxa-1-phosphatridecan-1-iumiodide (prepared as described in Example 27 step (a)) (1.99 g, 3.06mmol) in DCM (10 mL) was treated with TFA (2.5 mL, 32.4 mmol). Afterstirring at room temperature for 1 h the solvent was removed underreduced pressure. The resulting residue was co-evaporated with toluene(3×). The resulting residue was taken up in DCM (20 mL) and addeddropwise to a stirred solution of MTBE (200 mL). After 10 min theresulting gum was isolated by decantation of the MTBE and taken up inDCM (20 mL). The solvent was removed under reduced pressure to give12-carboxy-1,1,1-triphenyl-4,7,10-trioxa-1-phosphadodecan-1-ium iodide(1.8 g) as red gum, which was used in the next step without furtherpurification. LC-MS (Method G) 467 [M]⁺; RT 0.82 min

13-[(2R,3S,4R,5R,8R,10R,11R,12S,13S,14R)-11-{[(2S,3R,4S,6R)-4-(dimethylamino)-3-hydroxy-6-methyloxan-2-yl]oxy}-2-ethyl-3,4,10-trihydroxy-13-{[(2R,4R,5S,6S)-5-hydroxy-4-methoxy-4,6-dimethyloxan-2-yl]oxy}-3,5,8,10,12,14-hexamethyl-15-oxo-1-oxa-6-azacyclopentadecan-6-yl]-13-oxo-1,1,1-triphenyl-4,7,10-trioxa-1-phosphatridecan-1-iumchloride

Prepared following the procedure in Example 25 using2-carboxy-1,1,1-triphenyl-4,7,10-trioxa-1-phosphadodecan-1-ium iodide(prepared as described in Example 27 step (b)). Purification was bychromatography eluting with 0-15% MeOH/0.7 M NH₃ in DCM to give thetitle compound as a white solid.

LC-MS (Method H) 1184 [M]⁺; RT7.12 min

Example28—13-[(2R,3S,4R,5R,8R,10R,11R,12S,13S,14R)-11-{[(2S,3R,4S,6R)-4-(dimethylamino)-3-hydroxy-6-methyloxan-2-yl]oxy}-2-ethyl-3,4,10,13-tetrahydroxy-3,5,8,10,12,14-hexamethyl-15-oxo-1-oxa-6-azacyclopentadecan-6-yl]-13-oxo-1,1,1-triphenyl-4,7,10-trioxa-1-phosphatridecan-1-iumchloride

A solution of13-[(2R,3S,4R,5R,8R,10R,11R,12S,13S,14R)-11-{[(2S,3R,4S,6R)-4-(dimethylamino)-3-hydroxy-6-methyloxan-2-yl]oxy}-2-ethyl-3,4,10-trihydroxy-13-{[(2R,4R,5S,6S)-5-hydroxy-4-methoxy-4,6-dimethyloxan-2-yl]oxy}-3,5,8,10,12,14-hexamethyl-15-oxo-1-oxa-6-azacyclopentadecan-6-yl]-13-oxo-1,1,1-triphenyl-4,7,10-trioxa-1-phosphatridecan-1-iumchloride (prepared as described in Example 27) (55 mg, 0.05 mmol) inMeOH (1 mL) was treated with concentrated HCl (50 μL, 0.60 mmol). Theresulting reaction mixture was stirred at room temperature. After 1 hDCM (5 mL) was added and the organics washed with saturated aqueousNaHCO₃ (5 mL). The organic phase was passed through a phase separatorcontaining brine (10 mL) and the solvent removed under reduced pressure.The resulting residue was purified by silica column chromatographyeluting with 0-10% MeOH/0.7 M NH₃ in DCM to give a white solid, whichwas taken up in MeOH (3 mL) and filtered through an AmberliteIRA-400(Cl) ion exchange resin. The collected MeOH was recycled throughthe column 3×, followed by a fresh volume of MeOH. The combined MeOHwashings were concentrated under reduced pressure to give the titlecompound (18 mg) as a white solid.

LC-MS (Method H) 1026 [M]⁺; RT 6.57 min

Inhibition of Proliferation Assay

Cell Confluence

The confluence value is a surrogate for cell proliferation and growth.The value is expressed as a percent confluence, which represents thefraction of culture dish-surface that is occupied by cells. As thenumber of cells in the dish increases over time due to proliferation, sowill their coverage of that surface increase. Expansion of the cellpopulation on the cell culture-dish surface and confluence have mostly alinear relationship until the cells on the plate surface begin to reachsaturation or maximum density.

Confluence is determined based on image analysis. Image based softwarecan identify objects in the image field base on changes to pixel densityin a grey scale image. The software can then assign a mask to thosepixels within the object. Objects can be ‘gated’ out based on size andshape. To determine cell confluence, images of cells are first masked asobjects. The surface area of the image that is masked is measured andcompared to the total surface area of the culture dish surface to obtaina percent confluence.

MDA-231 cancer cells were obtained from ATCC. Cells were cultured inDulbecco's Modified Eagle Medium (DMEM) supplemented with 10% FetalBovine Serum (FBS), 2 mM Glutamax, 1 mM Non Essential Amino Acid (NEAA)solution and 1 mM sodium pyruvate. Compounds were dissolved in DMSO at10 mM and diluted in cellular medium and tested at 10 μM (micromolar).Final DMSO concentrations were <0.1%. Images were acquired with anIncuCyte Live Cell Imaging microscopy (Essen Bioscience) at every 3 hunder cell culture conditions with 10× objective over 4-5 d. Cellconfluence was calculated from one field of view per well using theIncuCyte in-built algorithm. Relative confluence values were obtained bynormalising each value to the time zero value in each sample.

Example Concentration uM % Confluence Azithromycin 100 17 Azithromycin10 99.1 1 10 4.2 2 10 13.4 3 10 98.4 4 10 89.9 5 10 5 6 10 118.9 7 1023.3 8 10 109.9 9 10 105.3 12 10 6.6 13 10 11 14 10 10.1 15 10 10.7 1610 9.9 17 10 82.6 18 10 9.6 19 10 9.7 20 10 13.5 21 10 8 22 10 102.7 2310 118 24 10 49.1 25 10 4.4

1. A compound comprising an ion of formula (I) or a pharmaceuticallyacceptable salt thereof:

wherein either Z¹ is

and Z² is R^(4b); or Z² is

and Z¹ is R^(2b); Z³ is independently selected from H, C(O)—C₁-C₆-alkylor Z³ has the structure:

Z⁴ is independently selected from H, C(O)—C₁-C₆-alkyl or Z⁴ has thestructure:

-L¹- is independently absent or is selected from —C(O)—, —C(O)O—,—S(O)₂—, —S(O)—, —C(O)NR⁵, and —S(O)₂NR⁵—; -L³- is independently at eachoccurrence either absent or selected from: —O—, —S—, —NR⁶—,—C(O)—,—OC(O)—, —C(O)O—, —S(O)₂—, —S(O)—, —NR⁵C(O)—, —C(O)NR⁵, —NR⁵S(O)₂—,—S(O)₂NR⁵—, —OC(O)NR⁵—,—NR⁵C(O)O—, NR⁵C(O)NR⁵, —CR⁷═CR⁷— and —C≡C—; -L²-and -L⁴- are each independently at each occurrence —C₁-C₄-alkylene-,each alkylene group being unsubstituted or substituted with from 1 to 6independently selected R⁸ groups; provided that any -L₂- or -L₄- groupthat is attached at each end to an atom selected from oxygen, nitrogen,sulphur or phosphorous is —C₂-C₄-alkylene-; -L⁵- is independently absentor is selected from —C(O)— and —C(O)NR⁵—; n is an integer selected from0, 1, 2, 3, 4 and 5; wherein L¹, L², L³, L⁴, L⁵ and n are selected suchthat length of the linker formed by those groups is from 3 to 16 atoms;R^(1a), R^(1b) and R^(1c) are each unsubstituted phenyl; R^(2a), R^(2b)and R^(2c) are each independently selected from H and C₁-C₆-alkyl;R^(3a) is independently selected from: H, C₁-C₆-alkyl andC(O)—C₁-C₆-alkyl; R^(3b) and R^(3c) are each independently selectedfrom: H and C(O)—C₁-C₆-alkyl; R^(4a) and R^(4b) are each independentlyselected from: H, C₁-C₆-alkyl and C(O)—C₁-C₆-alkyl; or R^(4a) and R^(4b)taken together form C(O); R⁵ is independently at each occurrenceselected from H and C₁-C₆-alkyl; R⁶ and R¹¹ are each independently ateach occurrence selected from: H, C₁-C₆-alkyl, C(O)C₁-C₆-alkyl andS(O)₂—C₁-C₆-alkyl; R⁷ is independently at each occurrence selected fromH, C₁-C₄-alkyl and halo; R⁸ is independently at each occurrence selectedfrom: C₁-C₆-alkyl, C₂-C₆-alkynyl, C₂-C₆-alkenyl, C₁-C₆-haloalkyl, OR⁹,SR¹⁰, NR¹⁰R¹¹, C(O)OR¹⁰, C(O)NR¹⁰R¹⁰, NR¹⁰C(O)OR¹⁰, OR¹⁰C(O)NR¹⁰R¹⁰,halo, cyano, nitro, C(O)R¹⁰, S(O)₂OR¹⁰, S(O)₂R¹⁰, S(O)R¹⁰ andS(O)₂NR¹⁰R¹⁰; R⁹ is independently at each occurrence selected from: H,C₁-C₆-alkyl and C₁-C₆-haloalkyl; R¹⁰ is independently at each occurrenceselected from: H and C₁-C₆-alkyl; and wherein any of the abovementionedalkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, heteroaryl orphenyl groups is optionally substituted where chemically allowable byfrom 1 to 4 groups independently selected from oxo, C₁-C₆-alkyl,C₂-C₆-alkynyl, C₂-C₆-alkenyl, C₁-C₆-haloalkyl, OR^(a), NR^(a)R^(b),SR^(a), C(O)OR^(a), C(O)NR^(a)R^(a), halo, cyano, nitro, C(O)R^(a),S(O)₂OR^(a), S(O)₂R^(a), S(O)R^(a) and S(O)₂NR^(a)R^(a); wherein R^(a)is independently at each occurrence selected from: H and C₁-C₆-alkyl;and R^(b) is independently at each occurrence selected from: H,C₁-C₆-alkyl, C(O)C₁-C₆-alkyl and S(O)₂—C₁-C₆-alkyl.
 2. A compound ofclaim 1, wherein Z¹ is

and Z² is R^(4b).
 3. A compound of claim 2, wherein L¹ is selected from—C(O)— and —S(O)₂—.
 4. A compound of claim 2, wherein L¹ is absent.
 5. Acompound of any one of claims 2 to 4, wherein R^(4a) is H and R^(4b) isH.
 6. A compound of any one of claims 2 to 4, wherein R^(4a) and R^(4b)together form C(O).
 7. A compound of claim 1, wherein Z² is

and Z¹ is R^(2b), preferably wherein R^(2b) is methyl.
 8. A compound ofclaim 7, wherein L⁵ is —C(O)—.
 9. A compound of claim 7, wherein L⁵ is—C(O)NR⁵—.
 10. A compound of any one of claims 7 to 9, wherein R^(4a) isH.
 11. A compound of any one of claims 1 to 10, wherein L³ is at eachoccurrence absent.
 12. A compound of any one of claims 1 to 11, whereinL¹, L², L³, L⁴, L⁵ and n are selected such that length of the linkerformed by those groups is from 8 to 14 atoms.
 13. A compound of any oneof claims 1 to 12, wherein Z³ is

preferably wherein R^(3c) is H.
 14. A compound of any one of claims 1 to13, wherein R^(2a) is methyl and R^(2c) is methyl.
 15. A compound of anyone of claims 1 to 14, wherein R^(3a) is H and R^(3b) is H.
 16. Acompound of any one of claims 1 to 14, wherein R^(3a) is H and R^(3b) isC(O)CH₃.
 17. A compound of claim 1, wherein the cation of formula (I) isselected from:


18. A compound of any one of claims 1 to 17, wherein the compound is formedical use.
 19. A compound of any one of claims 1 to 17, wherein thecompound is for use in the treatment of cancer.
 20. A method for thetreatment of cancer, wherein the method comprises the administration ofa therapeutically effective amount of a compound of any one of claims 1to
 17. 21. A pharmaceutical composition, wherein the compositioncomprises a compound of any one of claims 1 to 17 and one or morepharmaceutically acceptable excipients.